Tumor suppressor pathway in C. elegans

ABSTRACT

The invention provides novel lin-61 genes and polypeptides involved in cell fate determination and in cell proliferation, as well as methods for utilizing these genes, and their encoded polypeptides, in identifying compounds that modulate cell proliferation.

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority from U.S. provisionalapplication No. 60/208,802, filed on Jun. 2, 2000, which is herebyincorporated by reference.

STATEMENT AS TO FEDERALLY SPONSORED RESEARCH

[0002] The present research was supported by a grant from the NationalInstitutes of Health (Number GM 24663). The U.S. government has certainrights to this invention.

BACKGROUND OF THE INVENTION

[0003] The field of the invention is cell proliferation.

[0004] The nematode Caenorhabditis elegans (C. elegans) is well suitedfor developmental genetic studies because the entire cell lineage hasbeen mapped and is essentially invariant from one animal to the next.Thus, by comparing the cell lineage of a wild-type animal to that of amutant animal, the changes in cell fates caused by the mutation can bedetermined.

[0005] A number of mutations that alter cell lineage in C. elegans,termed lin mutations, were obtained in genetic screens conducted byHorvitz and Sulston in the late 1970's. A subset of the mutationsaffected the formation of the vulva, a structure on the ventral surfaceof C. elegans hermaphrodites through which eggs are laid and throughwhich sperm enters during cross-fertilization. Six vulval precursorcells have the potential to undertake a vulval cell lineage, as definedby the number and pattern of cell divisions. In a wild-type animal onlythree of these cells actually undertake vulval cell fates and thesethree cells generate the 22 cells that make up the adult vulva. Inmultivulva (Muv) animals, most or all of the six vulval precursor cellsundertake vulval cell fates. In addition to the cells required for theformation of a normal vulva, these mutant animals generate an excess ofcells which cause the formation of raised, vulva-like structures on theventral surface of the animal. On the other hand, a vulvaless (Vul)phenotype results when no or too few vulval precursor cells adopt vulvalcell fates.

[0006] Genetic and molecular analyses of Muv and Vul animals havedefined a Ras signal transduction pathway that mediates induction of thehermaphrodite vulva. This pathway includes the LIN-3 EGF-like ligand,the LET-23 receptor tyrosine kinase, the SEM-5 adaptor, LET-60 Ras, theKSR-1 kinase, LIN-45 Raf, MEK-2, and the MPK-1 MAP kinase, and regulatesthe activities of the ETS transcription factor LIN-1 and thewinged-helix transcription factor LIN-31 (reviewed by Horvitz andSternberg, Nature 351:535-41, 1991; Sundaram and Han, Bioessays18:473-480, 1996; Tan et al., Cell 93:569-580, 1998). Mutant animals inwhich this pathway is ectpically activated can display a Muv phenotype,whereas mutant animals that have reduced Ras pathway signaling candisplay a Vul phenotype.

[0007] The synthetic multivulva (synMuv) genes act in twofunctionally-redundant pathways as negative regulators of the nematodeRas signaling pathway. The first synthetic multivulva mutant wasidentified by Horvitz and Sulston. The two genetic loci mutated in thismutant were termed lin-8 and lin-9. Reduction-of-function mutations inboth of these loci were required for a multivulva phenotype. Subsequentgenetic screens identified a set of loci which fall into the same classas lin-8, termed class A genes, and genes which fall into the same classas lin-9, termed class B genes. In general, an animal with areduction-of-function mutation in any class A gene and areduction-of-function mutation in any class B gene will display amultivulva phenotype, while animals carrying one or more mutations ofthe same class have a wild-type vulval phenotype. These two classesappear to define two functionally redundant pathways that negativelyregulate the expression of vulval cell fates.

[0008] Thus far at least four class A loci (lin-8, lin-15A, lin-38, andlin-56) and at least fourteen class B loci (lin-9, lin-15B, lin-35,lin-36, lin-37, lin-13, lin-52, lin-53, lin-54, lin-55 (dpl-1), lin-61,hda-1, tam-1 (Hsieh et al., Genes & Dev. 13:2958-2970, 1999), and the C.elegans E2F1 homolog (efl-1)) have been identified genetically. lin-15encodes both A and B activities in two non-overlapping transcripts. Inaddition, lin-37, lin-35, lin-53, lin-52, lin-54, lin-55 (dpl-1),lin-15A, lin-15B, lin-36, lin-9, lin-55, and efl-1 have been cloned(Ceol and Horvitz, Molecular Cell 7:461-473, 2001; Clark et al.,Genetics 137:987-997, 1994; Huang et al., Mol. Biol. Cell 5:395-411,1994; Beitel et al., Gene 254:253-263, 2000; and PCT WO 98/54299).

[0009] A number of the synMuv family members encode polypeptides withsequence similarity to polypeptides involved in cancer development andprogression. For example, lin-35 encodes a homolog of the mammalianpocket protein family, which includes retinoblastoma protein (Rb), p107,and p130. This family of proteins has been the subject of intense studysince the cloning of Rb in 1986. Rb is a tumor suppressor gene;mutations that inactivate Rb predispose individuals to tumor formation.Most commonly, inactivation of Rb results in a type of eye cancer,retinoblastoma, although inactivating mutations in Rb have been found inother types of tumors. The Rb protein is thought to function as anegative regulator of cell cycle progression. A number of molecules thatinteract, both directly and indirectly, with Rb and the other pocketproteins have been characterized in mammalian cells.

[0010] Another synMuv family member, lin-53, encodes a homolog of p48, aprotein which has been shown to bind Rb. Although the functionalsignificance of the interaction between p48 and Rb is not fullyunderstood, recent studies suggest that p48 may play a role inremodeling chromatin structure. In addition, lin-55(dpl-1) encodes ahomolog of the DP family of proteins (Ceol and Horvitz, Molecular Cell7:461-473, 2001). DP family members, together with E2F proteins, bindDNA at specific sites, thereby regulating the transcription of genesthat are essential for cell cycle progression. Furthermore, pocketproteins such as Rb bind to the DP-E2F complex to repress transcription.

[0011] As in the nematode, Ras pathways have been found to control cellproliferation in a range of organisms from the yeast Saccharomycescerevisiae to humans. The Ras pathway defines one class of oncogenesignaling pathways; members of this pathway, most commonly Ras itself,have been shown to be mutated in a broad range of human cancers (Hunter,Cell 88:333-346, 1997). Accordingly, analysis of the Ras pathway, inparticular the vulval induction pathway, in C. elegans addresses thesignificant need of increasing our understanding of cancer in general.

SUMMARY OF THE INVENTION

[0012] We isolated and cloned three novel C. elegans genes, lin-8,lin-56, and lin-61, that are part of two synMuv pathways and wecharacterized several mutations in these genes. lin-8, lin-56, and lin61 genes, their mutants, and the proteins they encode, may be used ingenetic and biochemical model systems to further our understanding ofcell proliferative diseases, including cancer, as well as in diagnosingand treating cell proliferative diseases.

[0013] Accordingly, the first aspect of the invention features asubstantially pure nucleic acid encoding a LIN-8 polypeptide, where theLIN-8 polypeptide includes at least 130 contiguous amino acids of SEQ IDNO: 1 and modulates cell proliferation. In a preferred embodiment ofthis aspect of the invention, the amino acid sequence of the LIN-8polypeptide includes SEQ ID NO:1, and in another embodiment, the LIN-8polypeptide has an amino acid alteration relative to the sequence of SEQID NO: 1, for example, one that increases cell proliferation.

[0014] In other preferred embodiments of this aspect, the polynucleotidesequence of the nucleic acid includes SEQ ID NO:2, or at least 400contiguous nucleotides of SEQ ID NO:2. In addition, the polynucleotidesequence of the nucleic acid may include a mutant lin-8 nucleic acidsequence, for example, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ IDNO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ IDNO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ IDNO:42, SEQ ID NO:44, or SEQ ID NO:46.

[0015] A second aspect of the invention features a polypeptide having anamino acid sequence identical to any one of SEQ ID NO:17, SEQ ID NO:19,SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33,SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43,SEQ ID NO:45, or SEQ ID NO:47.

[0016] In a third aspect, the invention encompasses a substantially purenucleic acid encoding a LIN-56 polypeptide, where the LIN-56 polypeptideincludes at least 110 contiguous amino acids of SEQ ID NO:3 andmodulates cell proliferation. In addition, the amino acid sequence ofthe LIN-56 polypeptide may be SEQ ID NO:3. In a preferred embodiment ofthis aspect of the invention, the LIN-56 polypeptide has an amino acidalteration relative to the sequence of SEQ ID NO:3, for example, onethat increases cell proliferation.

[0017] In additional embodiments of this aspect of the invention, thepolynucleotide sequence of the nucleic acid includes SEQ ID NO:4, or atleast 400 contiguous nucleotides of SEQ ID NO:4. Furthermore, thepolynucleotide sequence of the nucleic acid may be a mutant lin-56nucleic acid sequence, such as SEQ ID NO:48.

[0018] A fourth aspect of the invention features a substantially purenucleic acid encoding a LIN-61 polypeptide, where the LIN-61 polypeptideincludes at least 130 contiguous amino acids of SEQ ID NO:5 andmodulates cell proliferation. In preferred embodiments of this aspect,the amino acid sequence of the LIN-61 polypeptide may be SEQ ID NO:5, orthe LIN-61 polypeptide has an amino acid alteration relative to thesequence of SEQ ID NO:5, for example, one that increases cellproliferation.

[0019] In another embodiment of this aspect of the invention, thepolynucleotide sequence of the nucleic acid may be SEQ ID NO:6, or itmay include at least 400 contiguous nucleotides of SEQ ID NO:6.Furthermore, the polynucleotide sequence of the nucleic acid may be amutant lin-61 nucleic acid sequence, for example, one having thesequence of SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, or SEQ ID NO:78.

[0020] A fifth aspect of the invention encompasses a polypeptide havingan amino acid sequence identical to any one of SEQ ID NO:70, SEQ IDNO:71, or SEQ ID NO:72.

[0021] In a sixth aspect, the invention features a vector including anucleic acid having a polynucleotide sequence, for example, SEQ ID NO:2,SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ IDNO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQID.NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:73, SEQ IDNO:74, or SEQ ID NO:75. Preferably, this vector is capable of directingexpression of the nucleic acid, for example, in a cell.

[0022] A seventh aspect of the invention encompasses a transgenic cellincluding a nucleic acid sequence encoding a lin-8, a lin-56, or alin-61 polypeptide, where the nucleic acid sequence is located in thegenome of the cell in a position in which it does not naturally occur.In addition, the nucleic acid sequence may be operably linked to aheterologous promoter.

[0023] Additional aspects of the invention feature purified antibodieswhich specifically bind to a LIN-8, LIN-56, or LIN-61 polypeptide.

[0024] In further aspects, the invention provides methods of modulatingproliferation of a cell which involve administering aproliferation-modulating amount of a polypeptide, or a nucleic acidencoding a polypeptide, having the amino acid sequence of SEQ ID NO: 1,SEQ ID NO:3, or SEQ ID NO:5 to a cell, for example, a mammalian cell,such as a human cell. In addition, the nucleic acid sequence may becontained in a vector.

[0025] In another aspect, the invention features a method of identifyinga compound that modulates cell proliferation, involving: (a) providing acell expressing a nucleic acid, for example, a lin-8, lin-56, or lin-61nucleic acid, or a reporter gene, operably linked to a lin-8, lin-56, orlin-61 promoter; (b) contacting the cell with a candidate compound; and(c) measuring the expression of the nucleic acid, where an alteration inthe level of expression of the nucleic acid indicates the presence of acompound that modulates cell proliferation. In preferred embodiments,step (c) involves measuring the expression of the protein encoded by thenucleic acid, for example by using an antibody that specifically bindsto a LIN-8, LIN-56, or LIN-61 polypeptide, or step (c) may also involvemeasuring the mRNA level of the nucleic acid.

[0026] An additional aspect of the invention provides a method ofidentifying a candidate compound, for example, a polypeptide, that bindsto a LIN-8, LIN-56, or LIN-61 polypeptide, involving: (a) providing thepolypeptide; (b) contacting the polypeptide with a candidate compound;and (c) measuring the binding of the candidate compound to thepolypeptide, where the binding indicates the presence of a candidatecompound that binds a LIN-8, LIN-56, or LIN-61 polypeptide.

[0027] Furthermore, in another aspect, the invention provides a methodof diagnosing an animal, for example, a mammal, such as a human, for thepresence of a cell proliferation disease, such as cancer, or for anincreased likelihood of developing a cell proliferation disease. Thismethod involves determining whether a nucleic acid sample obtained fromthe animal includes a mutant lin-8, lin-56, or lin-61 nucleic acid,where the presence of the mutant lin-8, lin-56, or lin-61 nucleic acidindicates that the animal has a cell proliferation disease, or is at anincreased likelihood of developing a cell proliferation disease. Inpreferred embodiments of this aspect of the invention, the mutant lin-8nucleic acid may be, for example, lin-8(n2738), lin-8(n2731),lin-8(n3606), lin-8(n3595), lin-8(n2739), lin-8(n3586), lin-8(n3588),lin-8(n111), lin-8(n2741), lin-8(n3585), lin-8(n3646), lin-8(n2376),lin-8(n2378), lin-8(n2403), lin-8(n2724), lin-8(n3607), lin-8(n3591),lin-8(n3609), or lin-8(n3581), the mutant lin-56 nucleic acid may be,for example, lin-56(n3355) or lin-56(n2728), and the mutant lin-61nucleic acid may be, for example, lin-61 (n3446), lin-61 (n3447), lin-61(n3624), or lin-61 (n3635).

[0028] In an additional aspect, the invention features a method ofdiagnosing an animal, for example, a mammal, such as a human, for thepresence of a cell proliferation disease, or an increased likelihood ofdeveloping a cell proliferation disease. This method involves measuringlin-8, lin-56, or lin-61 nucleic acid expression in a sample obtainedfrom the animal, where an alteration in the expression, relative to asample obtained from an unaffected animal, indicates that the animal hasa cell proliferation disease, or an increased likelihood of developing acell proliferation disease. In a preferred embodiment of this aspect,nucleic acid expression is measured by measuring the amount of theLIN-8, LIN-56, or LIN-61 polypeptide, for example, by using an antibodythat specifically binds to a LIN-8, LIN-56, or LIN-61 polypeptide in thesample. However, nucleic acid expression may also be measured bymeasuring the amount of lin-8, lin-56, or lin-61 mRNA in the sample.

[0029] In a final aspect, the invention provides a method of identifyinga nucleic acid that modulates cell proliferation. This method involves:(a) expressing in a cell (i) a first nucleic acid operably linked to afirst promoter, where the first promoter may be the lin-8, lin-56, orlin-61 promoter; and (ii) a second nucleic acid operably linked to asecond promoter; and (b) measuring the expression of the first nucleicacid, where a modulation in the expression of the first nucleic acid inthe presence of the second nucleic acid, indicates that the secondnucleic acid modulates cell proliferation. In a preferred embodiment ofthis aspect, the first nucleic acid is a lin-8, lin-56, or lin-61nucleic acid. Furthermore, the measuring in step (b) may also involvecomparing the amount of modulation in the expression of the firstnucleic acid seen in the presence of the second nucleic acid with thatseen in the presence of a control nucleic acid that does not modulatecell proliferation.

[0030] Definitions

[0031] By a “lin-8 nucleic acid” is meant a nucleic acid sequence, orfragment thereof, that is substantially identical to SEQ ID NO:2, orportions thereof. Preferably a “lin-8 nucleic acid” is identical to atleast 100, 200, 300, 390, 400, 450, 500, 600, 700, 800, 900, 1000, 1100,or 1200 contiguous nucleotides of SEQ ID NO:2, its complement, or to thecorresponding RNA sequence. However, a “lin-8 nucleic acid” may also beidentical to SEQ ID NO:2. In addition, a “lin-8 nucleic acid” may becharacterized by its ability to modulate cell proliferation.Furthermore, a “lin-8 nucleic acid” may refer to a nucleic acid sequenceincluding nucleic acids 375-989, 400-900, 450-850, 500-800, or 550-750of SEQ ID NO:2, or to a nucleic acid that hybridizes under highlystringent conditions to these regions of SEQ ID NO:2. For example,highly stringent conditions include hybridization at about 42° C. andabout 50% formamide, 0.1 mg/mL sheared salmon sperm DNA, 1% SDS, 2× SSC,10% Dextran sulfate, a first wash at about 65° C., about 2× SSC, and 1%SDS, followed by a second wash at about 65° C. and about 0.1× SSC.Alternatively, highly stringent conditions may include hybridization atabout 42° C. and about 50% formamide, 0.1 mg/mL sheared salmon spermDNA, 0.5% SDS, 5× SSPE, 1× Denhardt's, followed by two washes at roomtemperature and 2× SSC, 0.1% SDS, and two washes at between 55-60° C.and 0.2× SSC, 0.1% SDS. The terms “gene” and “nucleic acid sequence” maybe used interchangeably.

[0032] By a “mutant lin-8 nucleic acid” or a “mutated lin-8 nucleicacid” is meant a nucleic acid sequence, or fragment thereof, differingfrom the wild-type lin-8 nucleic acid sequence by at least onenucleotide. This nucleotide difference may result, for example, in the“mutant lin-8 nucleic acid” encoding a truncated LIN-8 protein or onecontaining a missense mutation. Preferably, the “mutant lin-8 nucleicacid” is identical to at least 100, 200, 300, 390, 400, 500, 600, 700,800, 900, or 1000 contiguous nucleotides of SEQ ID NO:2, its complement,or the corresponding mRNA sequence. Most preferably, the “mutant lin-8nucleic acid” is the lin-8(n2738), lin-8(n2731), lin-8(n3606),lin-8(n3595), lin-8(n2739), lin-8(n3586), lin-8(n3588), lin-8(n111),lin-8(n2741), lin-8(n3585), lin-8(n3646), lin-8(n2376), lin-8(n2378),lin-8(n2403), lin-8(n2724), lin-8(n3607), lin-8(n3591), lin-8(n3609), orlin-8(n3581) nucleic acid sequence. Furthermore, a C. elegans carrying amutant lin-8 nucleic acid sequence and a reduction of function mutationin any synMuv class B gene will display a multivulva phenotype.

[0033] By a “lin-56 nucleic acid” is meant a nucleic acid sequence, orfragment thereof, that is substantially identical to SEQ ID NO:4, orportions thereof. Preferably a “lin-56 nucleic acid” is identical to atleast 100, 200, 300, 330, 390, 400, 450, 500, 600, 700, 800, 900, or1000 contiguous nucleotides of SEQ ID NO:4, its complement, or to thecorresponding RNA sequence. However, a “lin-56 nucleic acid” may also beidentical to SEQ ID NO:4. In addition, a “lin-56 nucleic acid” may becharacterized by its ability to modulate cell proliferation.Furthermore, a “lin-56 nucleic acid” may refer to a nucleic acidsequence including nucleic acids 376-1108, 400-1000, 400-700, 450-950,450-800, 500-900, 550-850, or 600-800 of SEQ ID NO:4, or to a nucleicacid that hybridizes under highly stringent conditions to these regionsof SEQ ID NO:4. For example, highly stringent conditions includehybridization at about 42° C. and about 50% formamide, 0.1 mg/mL shearedsalmon sperm DNA, 1% SDS, 2× SSC, 10% Dextran sulfate, a first wash atabout 65° C., about 2× SSC, and 1% SDS, followed by a second wash atabout 65° C. and about 0.1× SSC. Alternatively, highly stringentconditions may include hybridization at about 42° C. and about 50%formamide, 0.1 mg/mL sheared salmon sperm DNA, 0.5% SDS, 5× SSPE, 1×Denhardt's, followed by two washes at room temperature and 2× SSC, 0.1%SDS, and two washes at between 55-60° C. and 0.2× SSC, 0.1% SDS.

[0034] By a “mutant lin-56 nucleic acid” or a “mutated lin-56 nucleicacid” is meant a nucleic acid sequence, or fragment thereof, differingfrom the wild-type lin-56 nucleic acid sequence by at least onenucleotide. This nucleotide difference may result, for example, in the“mutant lin-56 nucleic acid” encoding a truncated LIN-56 protein or onecontaining a missense mutation. Preferably, the “mutant lin-56 nucleicacid sequence” is identical to at least 100, 200, 330, 390, 400, 500,600, 700, 800, 900, or 1000 contiguous nucleotides of SEQ ID NO:4, itscomplement, or the corresponding mRNA sequence. Most preferably, the“mutant lin-56 nucleic acid” is the lin-56(n3355) nucleic acid sequence.Furthermore, a C. elegans carrying a mutant lin-56 nucleic acid sequenceand a reduction of function mutation in any synMuv class B gene willdisplay a multivulva phenotype.

[0035] By a “lin-61 nucleic acid” is meant a nucleic acid sequence, orfragment thereof, that is substantially identical to SEQ ID NO:6, orportions thereof. Preferably a “lin-61 nucleic acid” is identical to atleast 100, 200, 300, 390, 400, 450, 500, 600, 700, 800, 900, 1000, 1100,1200, 1300, or 1400 contiguous nucleotides of SEQ ID NO:6, itscomplement, or to the corresponding RNA sequence. However, a “lin-61nucleic acid” may also be identical to SEQ ID NO:6. In addition, a“lin-61 nucleic acid” may be characterized by its ability to modulatecell proliferation. Furthermore, a “lin-61 nucleic acid” may refer to anucleic acid sequence including nucleic acids 375-1150, 400-1100,400-1000, 450-950, 500-1000, 500-900, or 550-850 of SEQ ID NO:6, or to anucleic acid that hybridizes under highly stringent conditions to theseregions of SEQ ID NO:6. For example, highly stringent conditions includehybridization at about 42° C. and about 50% formamide, 0.1 mg/mL shearedsalmon sperm DNA, 1% SDS, 2× SSC, 10% Dextran sulfate, a first wash atabout 65° C., about 2× SSC, and 1% SDS, followed by a second wash atabout 65° C. and about 0.1× SSC. Alternatively, highly stringentconditions may include hybridization at about 42° C. and about 50%formamide, 0.1 mg/mL sheared salmon sperm DNA, 0.5% SDS, 5× SSPE, 1×Denhardt's, followed by two washes at room temperature and 2× SSC, 0.1%SDS, and two washes at between 55-60° C. and 0.2× SSC, 0.1% SDS.

[0036] However, a “lin-61 nucleic acid” may also be identical to atleast 100, 200, 300, 390, 450, 500, 600, 700, 800, 900, 1000, 1100,1200, 1300, or 1400 contiguous nucleotides of SEQ ID NO:76, itscomplement, or to the corresponding RNA sequence. In addition, a “lin-61nucleic acid” may be identical to SEQ ID NO:76.

[0037] By a “mutant lin-61 nucleic acid” or a “mutated lin-61 nucleicacid” is meant a nucleic acid sequence, or fragment thereof, differingfrom the wild-type lin-61 nucleic acid sequence by at least onenucleotide. This nucleotide difference may result, for example, in the“mutant lin-61 nucleic acid” encoding a truncated LIN-61 protein or onecontaining a missense mutation. Preferably, the “mutant lin-61 nucleicacid” is identical to at least 100, 200, 300, 390, 400, 500, 600, 700,800, 900, 1000, 1100, 1200, 1300, or 1400 contiguous nucleotides of SEQID NO:6, its complement, or the corresponding mRNA sequence. Mostpreferably, the “mutant lin-61 nucleic acid” is the lin-61(n3446),lin-61(n3447),or lin-61(n3624) nucleic acid sequence. However, a mutant“lin-61 nucleic acid,” for example, a lin-61(sy223) or lin-61(n3635)nucleic acid sequence, may also be identical to at least 100, 200, 300,390, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, or 1400contiguous nucleotides of SEQ ID NO:76, its complement, or thecorresponding mRNA sequence. Furthermore, a C. elegans carrying a mutantlin-61 nucleic acid sequence and a reduction of function mutation in anysynMuv class A gene will display a multivulva phenotype.

[0038] By a “lin-8(n111) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO: 16, its complement, or the corresponding RNA sequence.

[0039] By a “lin-8(n2741) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO: 18, its complement, or the corresponding RNA sequence.

[0040] By a “lin-8(n2738) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:20, its complement, or the corresponding RNA sequence.

[0041] By a “lin-8(n2731) nucleic. acid” is meant a nucleic acidsequence having SEQ ID NO:22, its complement, or the corresponding RNAsequence.

[0042] By a “lin-8(n3585) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:24, its complement, or the corresponding RNA sequence.

[0043] By a “lin-8(n3646) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:26, its complement, or the corresponding RNA sequence.

[0044] By a “lin-8(n3606) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:28, its complement, or the corresponding RNA sequence.

[0045] By a “lin-8(n2376) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:30, its complement, or the corresponding RNA sequence.

[0046] By a “lin-8(n2378) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:32, its complement, or the corresponding RNA sequence.

[0047] By a “lin-8(n3595) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:34, its complement, or the corresponding RNA sequence.

[0048] By a “lin-8(n2403) nucleic acid,” a “lin-8(2724) nucleic acid,”or a “lin-8(3607) nucleic acid” is meant a nucleic acid sequence havingSEQ ID NO:36, its complement, or the corresponding RNA sequence.

[0049] By a “lin-8(n3581) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:38, its complement, or the corresponding RNA sequence.

[0050] By a “lin-8(n3609) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:40, its complement, or the corresponding RNA sequence.

[0051] By a “lin-8(n2739) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:42, its complement, or the corresponding RNA sequence.

[0052] By a “lin-8(n3586) nucleic acid,” or a “lin-8(n3588) nucleicacid” is meant a nucleic acid sequence having SEQ ID NO:44, itscomplement, or the corresponding RNA sequence.

[0053] By a “lin-8(n3591) nucleic acid” is meant a nucleic acid sequencehaving SEQ ID NO:46, its complement, or the corresponding RNA sequence.

[0054] By a “lin-56(n3355) nucleic acid” is meant a nucleic acidsequence having SEQ ID NO:48, its complement, or the corresponding RNAsequence.

[0055] By a “lin-56(n2728) nucleic acid” is meant a deletionencompassing all or part of a lin-56 nucleic acid sequence in anisolated nucleic acid containing the genomic region that normallycontains a lin-56 nucleic acid sequence.

[0056] By a “lin-61 (n3446) nucleic acid” is meant a nucleic acidsequence having SEQ ID NO:73, its complement, or the corresponding RNAsequence.

[0057] By a “lin-61(n3447) nucleic acid” is meant a nucleic acidsequence having SEQ ID NO:74, its complement, or the corresponding RNAsequence.

[0058] By a “lin-61(n3624) nucleic acid” is meant a nucleic acidsequence having SEQ ID NO:75, its complement, or the corresponding RNAsequence.

[0059] By a “lin-61(sy223) nucleic acid” is meant a nucleic acidsequence having SEQ ID NO:77, its complement, or the corresponding RNAsequence.

[0060] By a “lin-61(n3635) nucleic acid” is meant a nucleic acidsequence having SEQ ID NO:78, its complement, or the corresponding RNAsequence.

[0061] By “LIN-8 polypeptide” or “LIN-8 protein” is meant a polypeptideor protein encoded by a lin-8 nucleic acid sequence. Preferably, a“LIN-8 polypeptide” is identical to at least 30, 50, 100, 130, 200, 250,300, or 350 contiguous amino acids of SEQ ID NO: 1. Most preferably, a“LIN-8 polypeptide” is identical to SEQ ID NO: 1 and has a LIN-8biological activity described below.

[0062] By a “mutant LIN-8 polypeptide” is meant a LIN-8 polypeptide thatdiffers by at least one amino acid from a wild-type LIN-8 polypeptide.In addition, a mutant LIN-8 polypeptide may also be a truncated protein,for example, due to the presence of a premature stop codon in thenucleic acid sequence encoding the mutant LIN-8 polypeptide. Inaddition, a “mutant LIN-8 polypeptide” is preferably 50%, 60%, 70%, 75%,80%, 85%, 90%, 95%, or even 99% identical to at least 30, 50, 100, 130,200, 250, 300, or 350 contiguous amino acids of SEQ ID NO:1.Furthermore, the mutant LIN-8 polypeptide preferably is the polypeptideencoded by lin-8(n2738), lin-8(n2731), lin-8(n3606), lin-8(n3595),lin-8(n2739), lin-8(n3586), lin-8(n3588), lin-8(n111), lin-8(n2741),lin-8(n3585), lin-8(n3646), lin-8(n2376), lin-8(n2378), lin-8(n2403),lin-8(n2724), lin-8(n3607), lin-8(n3591), lin-8(n3609), or lin-8(n3581).Most preferably, a “mutant LIN-8 polypeptide” is identical to SEQ IDNO:17, SEQ ID NO:19, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ IDNO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ IDNO:41, SEQ ID NO:43, SEQ ID NO:45, or SEQ ID NO:47.

[0063] By “LIN-56 polypeptide” or “LIN-56 protein” is meant apolypeptide or protein encoded by a lin-56 nucleic acid sequence.Preferably, a “LIN-56 polypeptide” is identical to at least 30, 50, 100,130, 200, 250, or 300 contiguous amino acids of SEQ ID NO:3. Mostpreferably, a “LIN-56 polypeptide” is identical to SEQ ID NO:3 and has aLIN-56 biological activity described below.

[0064] By a “mutant LIN-56 polypeptide” is meant a LIN-56 polypeptidethat differs by at least one amino acid from a wild-type LIN-56polypeptide. In addition, a mutant LIN-56 polypeptide may also be atruncated protein, for example, due to the presence of a premature stopcodon in the nucleic acid sequence encoding the mutant LIN-56polypeptide. In addition, a “mutant LIN-56 polypeptide” is preferably50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or even 99% identical to atleast 30, 50, 100, 130, 200, 250, or 300 contiguous amino acids SEQ IDNO:3. Furthermore, the mutant LIN-56 polypeptide is preferably thepolypeptide encoded by a lin-56(n3355) nucleic acid.

[0065] By “LIN-61 polypeptide” or “LIN-61 protein” is meant apolypeptide or protein encoded by a lin-61 nucleic acid sequence.Preferably, a “LIN-61 polypeptide” is identical to at least 30, 50, 100,130, 200, 250, 300, 350, or 400 contiguous amino acids of SEQ ID NO:5.Most preferably, a “LIN-61 polypeptide” is identical to SEQ ID NO:5 andhas a LIN-61 biological activity described below.

[0066] By a “mutant LIN-61 polypeptide” is meant a LIN-61 polypeptidethat differs by at least one amino acid from a wild-type LIN-61polypeptide. In addition, a mutant LIN-61 polypeptide may also be atruncated protein, for example, due to the presence of a premature stopcodon in the nucleic acid sequence encoding the mutant LIN-61polypeptide. In addition, a “mutant LIN-61 polypeptide” is preferably50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or even 99% identical to atleast 30, 50, 100, 130, 200, 250, 300, 350, or 400 contiguous aminoacids SEQ ID NO:5. Preferably, a “mutant LIN-61 polypeptide” is encodedby a lin-61(n3446), lin-61(n3447), or lin-61(3624) nucleic acid. Mostpreferably, a “mutant LIN-61 polypeptide” is identical to SEQ ID NO:70,SEQ ID NO:71, or SEQ ID NO:72.

[0067] By an “amino acid alteration,” as used herein, is meant a changein an amino acid sequence, relative to the wild-type sequence. Such achange may be, for example, the substitution of one or more amino acidswith heterologous amino acids, as well as the addition or deletion ofone or more amino acids. In addition, an “amino acid alteration” mayresult in a truncated protein.

[0068] By “heterologous promoter” is meant a nucleic acid sequence thatdrives expression of a nucleic acid sequence, e.g., a gene, with whichis not naturally associated.

[0069] By a “synMuv gene” is meant a nucleic acid sequence encodingLIN-9, LIN-15A, LIN-15B, LIN-37, LIN-35, LIN-53, LIN-55, LIN-52, LIN-54,and the E2F-1 gene of C. elegans, and the LIN-54 genes of the mouse andhuman. SynMuv genes also include those which encode polypeptides encodedby ESTs zp44h06.s1, zr79e11.r1, and EST180962 and any other nucleic acidsequence identified as a synMuv sequence known in the art.

[0070] By “synMuv polypeptide” or “synMuv protein” is meant apolypeptide encoded by a synMuv gene.

[0071] By “LIN-8 biological activity,” “LIN-56 biological activity,” or“LIN-61 biological activity” is meant an activity of a LIN-8, LIN-56, orLIN-61 polypeptide, when expressed or overexpressed, either alone or incombination, with another polypeptide in a cell, which is absent ordecreased in the absence of the LIN-8, LIN-56, or LIN-61 polypeptide. ALIN-8, LIN-56, or LIN-61 biological activity includes modulating oraltering cell proliferation. Another activity is rescuing (i.e.,suppressing) a LIN-8, LIN-56, or LIN-61 mutant phenotype. Lesspreferably, a LIN-8, LIN-56, or LIN-61 biological activity involvesbinding to other known synMuv polypeptides, in vivo or in vitro. AnotherLIN-8, LIN-56, or LIN-61 biological activity is binding to an antibodythat recognizes a LIN-8, LIN-56, or LIN-61 polypeptide. Finally, aLIN-8, LIN-56, or LIN-61 biological activity may also be the ability ofthe nucleic acid sequence encoding the polypeptide to hybridize to adetectably-labeled probe from a lin-8, lin-56, or lin-61 nucleic acidsequence under high, or less preferably, low stringency conditions.

[0072] By “modulating cell proliferation” or “altering cellproliferation” is meant increasing or decreasing the number of cellswhich undergo cell division in a given cell population or altering thefate of a given cell. It will be appreciated that the degree ofmodulation provided by LIN-8, LIN-56, LIN-61, or a modulatory compound,in a given assay will vary, but that one skilled in the art candetermine the statistically significant change (e.g., a p-value:≦0.05)in the level of cell proliferation which identifies a modulatorycompound.

[0073] By “inhibiting cell proliferation” is meant any decrease in thenumber of cells that undergo division relative to an untreated control.Preferably, the decrease is at least 25%, more preferably the decreaseis at least 50%, and most preferably the decrease is at least 75%, 80%,or even 100%.

[0074] By “a cell proliferation disease,” is meant a disorder that isdue to any genetic alteration within a differentiated cell that resultsin the abnormal proliferation of the cell. Examples of such changesinclude mutations in genes involved in the regulation of the cell cycle,of growth control, or of apoptosis, and further can include tumorsuppressor genes and proto-oncogenes. In addition, “a cell proliferationdisease” may be the result of, for example, an inappropriately highlevel of cell division, or an inappropriately low level of apoptosis, orboth. Specific examples of cell proliferation diseases are various typesof cancer including cancers of the reproductive system, such as cervicalcancer and ovarian cancer.

[0075] By “unaffected animal,” as used herein, is meant an animal thatdoes not have, or is not at an increased likelihood of developing, acell proliferation disease.

[0076] By “specifically binds,” as used herein in reference to anantibody, is meant an antibody that recognizes a specific protein, orshows staining in a sample, but does not recognize a specific protein,or show staining, in a sample not containing the protein of interest.Assays used to determine binding include, for example, Western blotting,affinity column purification, and tissue staining. A sample notcontaining the protein of interest may be obtained from an organismmutant for the protein of interest.

[0077] By “polypeptide” is meant any chain of amino acids, regardless oflength or post-translational modification (e.g., glycosylation orphosphorylation).

[0078] By “substantially identical” is meant a polypeptide or nucleicacid exhibiting at least 50%, preferably 60%, 70%, 75%, or 80%, morepreferably 90% or 95%, and most preferably 99% homology to a referenceamino acid or nucleic acid sequence. For polypeptides, the length ofcomparison sequences will generally be at least 16 amino acids,preferably at least 20 amino acids, more preferably at least 25, 35, 50,75, 100, 110, 130, 150, 200, 250, 300, or 310 amino acids, and mostpreferably the full-length amino acid sequence. For nucleic acids, thelength of comparison sequences will generally be at least 50nucleotides, preferably at least 60 nucleotides, more preferably atleast 75, 110, 200, 330, 390, 450, 600, 800, 900, or 1000 nucleotides,and most preferably the full-length nucleotide sequence.

[0079] Sequence identity may be measured using sequence analysissoftware on the default setting (i.e., Sequence Analysis SoftwarePackage of the Genetics Computer Group, University of WisconsinBiotechnology Center, 1710 University Avenue, Madison, Wis. 53705). Suchsoftware may match similar sequences by assigning degrees of homology tovarious substitutions, deletions, and other modifications. Conservativesubstitutions typically include substitutions within the followinggroups: glycine, alanine, valine, isoleucine, leucine; aspartic acid,glutamic acid, asparagine, glutamine; serine, threonine; lysine,arginine; and phenylalanine, tyrosine.

[0080] A polypeptide which is substantially identical to a LIN-8,LIN-56, or LIN-61 polypeptide (SEQ ID NOS:1, 3, or 5) may be, forexample, another substantially pure naturally-occurring mammalian LIN-8,LIN-56, or LIN-61 polypeptide as well as an allelic variant; a naturalmutant; an induced mutant; and a DNA sequence that encodes apolypeptide. In addition, such polypeptides may also be any polypeptidesspecifically bound by antisera directed to a LIN-8, LIN-56, or LIN-61polypeptide. Furthermore, polypeptides that are substantially identicalto LIN-8, LIN-56, or LIN-61 polypeptides also include chimericpolypeptides that have a LIN-8, LIN-56, or LIN-61 polypeptide portion.Preferably, this LIN-8, LIN-56, or LIN-61 polypeptide portion containsat least 50, 75, 90, 110, 130, 150, 200, 250, or 300 contiguous aminoacids of SEQ ID NOS: 1, 3, or 5.

[0081] By a “substantially pure polypeptide” is meant a polypeptide, forexample, LIN-8, LIN-56, or LIN-61, that has been separated fromcomponents which naturally accompany it. Typically, the polypeptide issubstantially pure when it is at least 60%, by weight, free from theproteins and naturally-occurring organic molecules with which it isnaturally associated. Preferably, the preparation is at least 75%, morepreferably at least 90%, and most preferably at least 99%, by weight, aLIN-8, LIN-56, or LIN-61 polypeptide. A substantially pure LIN-8,LIN-56, or LIN-61 polypeptide may be obtained, for example, byextraction from a natural source (e.g., a fibroblast, neuronal cell, orlymphocyte cell); by expression of a recombinant nucleic acid encoding aLIN-8, LIN-56, or LIN-61 polypeptide; or by chemically synthesizing thepolypeptide. Purity can be measured by any appropriate method, e.g.,column chromatography, polyacrylamide gel electrophoresis, or by HPLCanalysis.

[0082] A protein is substantially free of naturally associatedcomponents when it is separated from those contaminants, which accompanyit in its natural state. Thus, a protein, which is chemicallysynthesized or produced in a cellular system different from the cellfrom which it naturally originates, will be substantially free from itsnaturally associated components. Accordingly, substantially purepolypeptides include those derived from eukaryotic organisms butsynthesized in E. coli or other prokaryotes.

[0083] By a “substantially pure DNA” or “a substantially pure nucleicacid sequence” is meant a nucleic acid sequence or DNA that is free ofthe genes which, in the naturally-occurring genome of the organism fromwhich the nucleic acid sequence or DNA of the invention is derived,flank the gene. The term therefore includes, for example, a recombinantDNA which is incorporated into a vector; into an autonomouslyreplicating plasmid or virus; or into the genomic DNA of a prokaryote oreukaryote; or which exists as a separate molecule (e.g., a cDNA or agenomic or cDNA fragment produced by PCR or restriction endonucleasedigestion) independent of other sequences. It also includes arecombinant DNA which is part of a hybrid gene encoding additionalpolypeptide sequence or an antisense DNA or RNA sequence.

[0084] By “antisense” is meant a nucleic acid sequence, regardless oflength, that is complementary to the coding strand gene encoding anucleic acid sequence of interest, for example, a lin-8, lin-56, orlin-61 nucleic acid sequence. Preferably, the antisense nucleic acid iscapable of decreasing the biological activity of the polypeptide encodedby the nucleic acid sequence of interest when present in a cell.Preferably the decrease is at least 10%, relative to a control, morepreferably 25%, 50%, or 75%, and most preferably 100%.

[0085] By “analog of LIN-8”, “analog of LIN-56”, or “analog of LIN-61,”is meant differing from a naturally-occurring LIN-8, LIN-56, or LIN-61polypeptide by amino acid sequence differences, by post-translationalmodifications, or by both. Analogs of the invention will generallyexhibit at least 85%, more preferably 90%, and most preferably 95% oreven 99% identity with all or part of a naturally-occurring LIN-8,LIN-56, or LIN-61 amino acid sequence (SEQ ID NOS: 1, 3, or 5). Thelength of sequence comparison is at least 25, 50, 75, 100, 110, 130,150, 200, 250, or 300 contiguous amino acid residues, and morepreferably more than 310 amino acid residues, for example, thefull-length sequence. Modifications include in vivo and in vitrochemical derivatization of polypeptides, e.g., acetylation,carboxylation, phosphorylation, or glycosylation; such modifications mayoccur during polypeptide synthesis or processing or following treatmentwith isolated modifying enzymes.

[0086] In addition, analogs may differ from the naturally-occurringLIN-8, LIN-56, or LIN-61 polypeptide by alterations in primary sequence.These include genetic variants, both natural and induced (for example,resulting from random mutagenesis by irradiation or exposure toethanemethylsulfate (EMS) or by site-specific mutagenesis as describedin Sambrook, Fritsch and Maniatis, Molecular Cloning: A LaboratoryManual (2d ed.), Cold Spring Harbor Press, 1989, or Ausubel et al.,Current Protocols in Molecular Biology, Wiley Interscience, New York,2000). Also included are cyclized peptides, molecules, and analogs whichcontain residues other than L-amino acids, e.g., D-amino acids ornon-naturally-occurring or synthetic amino acids, e.g., β or γ aminoacids.

[0087] As used herein, the term “LIN-8 polypeptide fragment”, “LIN-56polypeptide fragment”, or “LIN-61 polypeptide fragment,” means at least20 contiguous amino acids, preferably at least 50 contiguous aminoacids, more preferably at least 100 contiguous amino acids, and mostpreferably at least 110, 130, 150, 200, 250, 300, 310 or more contiguousamino acids of SEQ ID NOS: 1, 3, or 5. Fragments of LIN-8, LIN-56, orLIN-61 polypeptides can be generated by methods known to those skilledin the art or may result from normal protein processing (e.g., removalof amino acids from the nascent polypeptide that are not required forbiological activity or removal of amino acids by alternative mRNAsplicing or alternative protein processing events). Preferable fragmentsor analogs according to the invention are those which facilitatespecific detection of a lin-8, lin-56, or lin-61 nucleic acid or aminoacid sequence in a sample to be diagnosed.

[0088] By “transformed cell” is meant a cell into which (or into anancestor of which) has been introduced, by means of recombinant DNAtechniques, a DNA molecule encoding (as used herein) a LIN-8, LIN-56,LIN-61 polypeptide, or a reporter gene.

[0089] By “transgene” is meant any piece of DNA that is inserted byartifice into a cell, and becomes part of the genome of the organismthat develops from that cell. Such a transgene may include a gene thatis partly or entirely heterologous (i.e., foreign) to the transgenicorganism, or may represent a gene homologous to an endogenous gene ofthe organism. Self-replicating units, such as artificial chromosomes,are included.

[0090] By “transgenic” is meant any cell that includes a DNA sequenceinserted by artifice into a cell and that becomes part of the genome ofthe organism that develops from that cell. As used herein, thetransgenic organism is generally a transgenic nematode (e.g., C.elegans), non-human mammal (e.g., a rodent such as a rat or mouse), or aplant, and the DNA sequence (transgene) is inserted by artifice into thenuclear genome.

[0091] By “transformation” is meant any method for introducing foreignmolecules into a cell. Microinjection, lipofection, calcium phosphateprecipitation, retroviral delivery, electroporation, and biolistictransformation are just a few of the teachings which may be used. Forexample, biolistic transformation is a method for introducing foreignmolecules into a cell using velocity driven microprojectiles such astungsten or gold particles. Such velocity-driven methods originate frompressure bursts, which include, but are not limited to, helium-driven,air-driven, and gunpowder-driven techniques. Biolistic transformationmay be applied to the transformation or transfection of a wide varietyof cell types and intact tissues including, without limitation,intracellular organelles, bacteria, yeast, fingi, algae, animal tissue,and cultured cells.

[0092] By “positioned for expression” is meant that a nucleic acidmolecule is positioned adjacent to a nucleic acid sequence that directstranscription and translation of the sequence (i.e., facilitates theproduction of, e.g., a LIN-8, LIN-56, or LIN-61 polypeptide, arecombinant protein or an RNA molecule).

[0093] By “reporter gene” is meant a gene whose expression may beassayed; such genes include, without limitation, those encodingglucuronidase (GUS), luciferase, chloramphenicol transacetylase (CAT),green fluorescent protein (GFP), and β-galactosidase.

[0094] By “promoter” is meant minimal sequence sufficient to directtranscription. Also included in the invention are those promoterelements that are sufficient to render promoter-dependent geneexpression controllable for specific cell-types or tissues. In addition,such promoters may also render gene expression inducible by externalsignals or agents. These promoter elements may be located in the 5′ or3′ regions of the native gene, for example, a lin-8, lin-56, or lin-61gene.

[0095] By “operably linked” is meant that a gene and a regulatorysequence(s) are connected in such a way as to permit gene expressionwhen the appropriate molecules (e.g., transcriptional activatorproteins) are bound to the regulatory sequence(s).

[0096] By “detectably-labeled” is meant any means for marking andidentifying the presence of a molecule, e.g., an oligonucleotide probeor primer, a gene or fragment thereof, or a cDNA molecule. Methods fordetectably-labeling a molecule are well known in the art and include,without limitation, radioactive labeling (e.g., with an isotope such as³²P or ³⁵S) and nonradioactive labeling (e.g., chemiluminescentlabeling, such as fluorescein labeling).

[0097] By “purified antibody” is meant an antibody that is at least 60%,by weight, free from proteins and naturally-occurring organic moleculeswith which it is naturally associated. Preferably, the preparation is atleast 75%, more preferably 80%, 85%, or 90%, and most preferably atleast 99%, by weight, antibody, e.g., a LIN-8, LIN-56, or LIN-61specific antibody. A purified antibody may be obtained, for example, byaffinity chromatography using recombinantly-produced protein orconserved motif peptides and standard techniques.

[0098] By “specifically binds” is meant an antibody which recognizes andbinds a protein, but which does not substantially recognize and bindother molecules in a sample, e.g., a biological sample, which naturallyincludes additional proteins.

[0099] By a “candidate compound” or “test compound” is meant a chemical,be it naturally-occurring or artificially-derived, that is surveyed forits ability to modulate cell proliferation, by employing one of theassay methods described herein. Candidate compounds may include, forexample, peptides, polypeptides, synthetic organic molecules,naturally-occurring organic molecules, nucleic acid molecules, andcomponents thereof.

[0100] Other features and advantages of the invention will be apparentfrom the following description of the preferred embodiments thereof, andfrom the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

[0101]FIG. 1 is a schematic representation of the mapping of lin-8.

[0102]FIG. 2 is a schematic representation of the rescue of lin-8 byB0454.1.

[0103]FIG. 3 shows the sequence homology of LIN-8 (SEQ ID NO:1) withseveral other polypeptides in C. elegans (SEQ ID NOS:55-67).

[0104]FIG. 4 is a schematic representation of the mapping of lin-56.

[0105]FIG. 5 is a schematic representation of the rescue of lin-56 byZK673.3.

[0106]FIG. 6 shows the LIN-56 polypeptide sequence (SEQ ID NO:3) as wellas an alignment indicating the homology of an internal region of LIN-56(SEQ ID NO:51) with several other C. elegans polypeptides (SEQ IDNOS:52-54).

[0107]FIG. 7 shows the sequence homology of the mbt repeats present inLIN-61 (SEQ ID NO:5) compared with mbt repeats from transcriptionalrepressors in other species (SEQ ID NOS:7-15).

[0108]FIG. 8 shows a sequence alignment between LIN-61 (SEQ ID NO:5) andpredicted worm (SEQ ID NO:69) and human (SEQ ID NO:68) proteins ofunknown function.

[0109]FIG. 9 shows that LIN-56 is localized to the nuclei of wild-typeC. elegans embryos (Panels A-C), larvae (Panel D), and adults. Thestaining shown was obtained with affinity-purified and pre-adsorbedrabbit polyclonal antibody HM1923.

[0110]FIG. 10 shows that LIN-56 staining is absent in lin-56(n2728)embryos (Panels A-C), larvae (Panel D), and adults. The staining shownwas obtained with affinity-purified and pre-adsorbed rabbit polyclonalantibody HM1923.

[0111]FIG. 11 shows that lin-61(RNAi) embryos display a failure inchromosome condensation during the first abortive mitotic division.

[0112]FIG. 12 shows that lin-61(RNAi) embryos display early embryoniclethality and a failure to complete cytokineses.

[0113]FIG. 13 shows that lin-61(RNAi) embryos arrest as multiplynucleated single cytoplasms.

DETAILED DESCRIPTION OF THE INVENTION

[0114] 1. Introduction

[0115] We have cloned lin-8, lin-56, and lin-61 of the C. elegans synMuvgene family. lin-8 and lin-56 are class A genes, and lin-61 is a class Bgene. These genes function in cell proliferation and are members of atumor suppressor pathway, which is related to a tumor suppressor pathwayof clinical importance in humans. Accordingly, the genes describedherein, mutations in these genes, as well as the previously known synMuvgenes, may be used to identify new tumor suppressors in other species,such as mammals, and may be used to identify therapeutic compounds. Theencoded polypeptides, and nematodes carrying the newly cloned genes ormutations in these genes may similarly be employed. lin-8 encodes anovel polypeptide. The invention provides the polypeptide sequence (SEQID NO: 1), the nucleic acid sequence (SEQ ID NO: 2), and lin-8 mutants,for example, lin-8(n2738), lin-8(n2731), lin-8(n3606), lin-8(n3595),lin-8(n2739), lin-8(n3586), lin-8(n3588), lin-8(n111), lin-8(n2741),lin-8(n3585), lin-8(n3646), lin-8(n2376), lin-8(n2378), lin-8(n2403),lin-8(n2724), lin-8(n3607), lin-8(n3591), lin-8(n3609) and lin-8(n3581).

[0116] We also cloned lin-56 and describe the LIN-56 polypeptidesequence (SEQ ID NO: 3) and the lin-56 nucleic acid sequence (SEQ ID NO:4) in the present invention, as well as lin-56 mutants, such aslin-56(n3355) and lin-56(n2728).

[0117] In addition, we cloned lin-61 (SEQ ID NO:6). The polypeptideencoded by this gene (SEQ ID NO:5) has homology with C. elegans andhuman polypeptides of unknown function, and to Drosophila polycomb groupmembers and their related human proteins. Furthermore, we identifiedlin-61 mutants, for example, lin-61(n3446), lin-61(n3447),lin-61(n3624), and lin-61(n3635).

[0118] The present invention also provides mammalian homologs of thenovel lin-8, lin-56, or lin-61 genes, which may be obtained usingroutine methods known to those skilled in the art. Such homologs mayfunction in activating, enhancing, or otherwise intensifying the effectsof tumor suppressors or oncogenes in mammals.

[0119] Genetic enhancer or suppressor screens may be performed toidentify new genes that may function in initiating, enhancing, orotherwise interfacing with this tumor suppressor pathway. In addition,the identification of the lin-8, lin-56, or lin-61 genes, in combinationwith what is known about proliferative disease pathways in mammals,allows one skilled in the art to readily devise drug screens involvingthese genes to search for compounds that affect cell proliferation.Specifically, compounds that block the Muv phenotype of animals with amutation in lin-8, lin-56, or lin-61 mutant animals are potentialanti-tumor agents. The Muv phenotype may be present, for example, in aC. elegans with either a mutation in lin-8 or lin-56 in combination witha reduction of function mutation in a synMuv B gene, or a mutation inlin-61 in combination with a reduction of function mutation in a synMuvA gene. Compounds that stimulate cell division in animals with a single,silent lin-8, lin-56, or lin-61 mutation are likely to be agonists ofcell proliferation and may act in a manner analogous to growth factors.

[0120] By providing insight regarding the function of the lin1-8,lin-56, or lin-61 members of the synMuv genes in tumor suppression, andby identifying mutants in these genes, we have provided, in concert withgenerally known molecular biology and nematode genetic methods, thenecessary elements of such methods and the compounds required for thepractice of such methods.

[0121] 2. Cloning of lin-8

[0122] The lin-8(n111) allele was isolated in an EMS screen forcell-lineage mutants (Horvitz and Sulston, Genetics 96:435-454, 1980).The lin-8 gene was mapped to the eight-map-unit interval between sup-9and lin-31 on chromosome II (Ferguson and Horvitz, Genetics 123:109-121,1989). As there were no other cloned genes in this interval that couldbe used for finer mapping, we used deficiencies to more precisely locatelin-8 on the physical map. The left endpoints of three deletions,ccDf112, ccDf1, and ccDf2, that remove lin-31 but not sup-9 hadpreviously been roughly located and we further mapped the left endpointof ccDf1 using PCR techniques. Analysis of the phenotypes of the Dfheterozygotes revealed that lin-8 is only deleted by ccDf11, thusplacing lin-8 between sup-9 and M151 (FIG. 1).

[0123] Further mapping against the polymorphism pPK5363 placed lin-8between sup-9 and C17F4. We identified a pool of two cosmids in thisregion that rescued the lin-8(lf) synMuv phenotype in germlinetransformation experiments. We further determined that this rescueeffect was attributable to the single cosmid C03E12. Germlinetransformation experiments with subclones of C03E12 indicated that lin-8is encoded by the predicted gene B0454.1 (FIG. 2). RNAi of this openreading frame produces a class A synMuv phenotype. Furthermore, nineteenalleles of lin-8 have been sequenced and all alleles contain mutationswithin B0454.1 (Table 1). TABLE 1 Mutations Identified in lin-8 AllelesMutant Amino lin-8 allele WT sequence Sequence Acid Change n2738 TGG TAGW79amber n2731 CAA TAA Q113ochre n3606 TGG TGA W147opal n3595 TGG TAGW163amber n3609 CAG TAG Q279amber n2739 AGA TGA R304opal n3586, n3588CAA TAA Q340ochre n111 CTG CCG L20P n2741 GTG ATG V68M n3585 CGC CACR127H n3646 CGC CAC R146H n2376 GAG AAG E148K n2378 CGC TGC R154C n2403,n2724, GAG AAG E164K n3607 n3591 GAG AAG E347K n3581 GTG -TG frameshiftafter aa192

[0124] The LIN-8 polypeptide is 386 amino acids in length, is novel, andappears to be highly charged. However, the LIN-8 polypeptide sharessequence homology with several other C. elegans polypeptides (FIG. 3).

[0125] 3. Cloning of lin-56

[0126] Identified as part of a screen to isolate new synMuv class Agenes, lin-56 was previously mapped to the three-map-unit intervalbetween dpy-10 and unc-53, close to unc-4, on chromosome II. We used thenumerous deficiencies available in this region to further delineate thephysical position of lin-56. The phenotypes of the Df heterozygotessuggested that lin-56 is positioned between the cloned markers stP98 andbli-1 (FIG. 4). We identified a pool of five cosmids in this region thatrescued the lin-56(lf) synMuv phenotype in germline transformationexperiments using a lin-56(n2728); lin-15B(n744) strain. This rescueeffect was attributed to the single cosmid ZK673 (FIG. 5).

[0127] Further germline transformation experiments with ZK673 subcloneslimited the lin-56 candidates to two predicted genes, ZK673.3 andZK673.4. Using a combination of Southern blotting and PCR techniques, wedetermined that lin-56(n2728) contains an 11.2 kb deletion thatcompletely eliminates these two genes and also removes the 3′ end of athird gene, ZK673.2. RNAi of ZK673.3, but not ZK673.4, produced a synMuvphenotype in a lin-15B(n744) background.

[0128] We have also identified a second lin-56 allele, designatedlin-56(n3355), isolated in a lin-56(n2728) non-complementation screen.Sequencing of this gene revealed that lin-56(n3355) contains a stopcodon in ZK673.3, confirming our identification of this open readingframe as lin-56.

[0129] The LIN-56 polypeptide is 322 amino acids in length (SEQ IDNO:3). This polypeptide appears to be novel and highly charged. LIN-56does have an internal sequence (SEQ ID NO:51) that shares weaksimilarities with sequences in ZK673.4, LIN-15A, and a predictedpolypeptide T25B9.8 (SEQ ID NOS:52-54)(FIG. 6). This region (SEQ IDNO:5 1) contains a C3H motif, which consists of a series of threecysteines followed by a histidine(C-x(2)-C-x(16)-A-x(7)-V-x(9)-A-x(11)-C-x(2)-H), where “x” can be anyamino acid and where the number in parentheses indicates the number of“x” amino acids present at that position of the motif. The C3H motifsuggests the presence of a metal binding domain, for example, azinc-finger domain. However, the spacing of these residues is unlikethat seen in any of the traditional motifs of this type.

[0130] In addition, antibodies to LIN-56 have been generated, using thefull length LIN-56 polypeptide to form a fusion protein with glutathioneS-transferase. These antibodies were purified against a fusion betweenfull length LIN-56 and maltose binding protein (MBP). The antibodiesrecognize the LIN-56 polypeptide in nematode extracts, as assessed byWestern blot analysis.

[0131] 4. Cloning of lin-61

[0132] We cloned the class B synthetic multivulva gene lin-61 by meansof mapping, transformation rescue with cosmid R06C7, and the directsequencing of multiple mutant alleles (see below). The cDNA sequence wasdetermined based on the intron/exon structure of the genomic DNA. Thepredicted cDNA contains an SL1 splice leader sequence. Based upon acombination of sequence from the 5′ and 3′ ends of the cDNAs, and thegenomic sequence of predicted ORF R06C7.7, we found lin-61 to encode aprotein highly similar to predicted human and C. elegans proteins ofundetermined function (FIG. 8).

[0133] The predicted LIN-61 protein contains motifs associated withtranscriptional repressor proteins in species ranging from Drosophila tohuman, indicating that LIN-61 may play a similar role in transcriptionalrepression. The predicted LIN-61 protein contains four mbt repeats.These motifs are present in members of the Drosophila melanogasterpolycomb family of transcriptional repressor proteins, including theproteins lethal(3) malignant brain tumor (1(3)mbt), and sex combs on themidleg (SCM). These motifs are also present in homologs of polycombfamily members in other species, including human (FIG. 7). Analysis ofmutant SCM alleles in Drosophila suggests that these mbt repeats areessential for SCM function (Bowermann et al., Genetics 150:675-686,1998).

[0134] In addition, we found lin-61 to be 41.1% identical at thenucleotide level to Drosophila 1(3)mbt, and to be 40.8% identical at thenucleotide level to the human 1(3)mbt homolog. Homology at the proteinlevel, however, is much greater within the mbt repeats.

[0135] We have isolated five alleles of lin-61 in addition to themutation sy223 that originally defined the gene. A total of five alleles(including sy223) have been sequenced in our lab. sy223 is a single basepair alteration (G to A at position 2228 of SEQ ID NO:76) altering thesplice acceptor site prior to the final exon of lin-61. lin-61(n3446) issingle base pair alteration (C to T at position 1234 of SEQ ID NO:6)causing an in frame ochre stop codon in place of Glutamine 413 of SEQ IDNO:5 and lin-61(n3447) is a single base pair alteration (G to A atposition 1061 of SEQ ID NO:6) causing an Serine to Asparagine missensemutation at amino acid 355 of SEQ ID NO:5. Furthermore, we identifiedlin-61(n3624) to be a single base pair alteration (C to T at position394 of SEQ ID NO:6) that causes a Proline to Serine missense mutation atamino acid 133 of SEQ ID NO:5 and lin-61(n3635) to encompass a singlebase pair alteration (G to A at position 1137 of SEQ ID NO:76) in thesplice acceptor site prior to the putative fourth exon of the protein.

[0136] Each of these alleles causes a synthetic multivulva phenotype incombination with a class A synMuv gene. This is the only phenotype wehave discovered to be associated with these mutations, and the allelelin-61(n3446) has wild-type vulval morphology when not in combinationwith a class A synMuv gene.

[0137] None of the lin-61 alleles described above represent a clearmolecular null. We have therefore sought to determine the phenotypeassociated with complete loss of lin-61 function by means ofRNA-mediated gene interference (RNAi). When wild-type animals or animalscontaining a mutation in a class A synMuv gene are injected with dsRNAsynthesized from cDNA corresponding to a portion of lin-61, they produceprogeny which suffer from completely penetrant embryonic lethality. Thisphenotype includes an inability to complete cytokinesis beginning at thesingle cell stage (FIG. 12A). In these embryos, the cytokinetic furrowforms and, at first, ingresses essentially normally (FIG. 12B-D), butsubsequently regresses (FIG. 12E-G), resulting in a polyploid singlecytoplasm (FIG. 12H). DNA replication continues to occur in the absenceof completed cytokinesis, and the embryos display a terminal phenotypein which high levels of DNA (as visualized by DAPI staining (FIG. 13B))and multiple mitotic spindles (as visualized by anti-tubulin antibodystaining (FIG. 13C)) are present within what is often a single cytoplasm(FIG. 13A). Moreover, embryos from lin-61(RNAi) mothers (FIG. 11A)display a failure to condense mitotic chromosomes (as visualized by DAPIstaining (FIG. 11B)). In these embryos, the mitotic spindle wasvisualized with an anti-tubulin antibody (FIG. 11C).

[0138] We further examined the developmental functions of lin-61 byinjecting lin-61 dsRNA into RNAi defective rde-1 hermaphrodites and thenmating these animals with N2 males (a technique described by Herman,Development 128:581-590, 2001). Cross progeny were then observed in aneffort to gain an understanding of the phenotype produced after thereduction of zygotic lin-61 activity, while maintaining at least somematernal lin-61 function. Unlike lin-61(RNAi), this approach yieldsanimals that reach adulthood but display a host of developmentalabnormalities including uncoordinated movement, abnormal development ofthe male tail, and possible abnormalities in the structure of the vulva.

[0139] Based on these results we conclude that lin-61 functions in avariety of embryonic and post-embryonic developmental processes inaddition to its role in vulval development and cell proliferation.

[0140] 5. Characterization of Interactions among the lin-8. lin-56,lin-61 and Other synMuv Gene Products

[0141] Standard yeast two-hybrid techniques are used to characterize thephysical interactions between the lin-8, lin-56, lin-61 and other synMuvgene products, for example, as described in U.S. Ser. No. 09/220,091,incorporated herein by reference. These two-hybrid systems can also beused to detect therapeutic compounds, which disrupt the synMuvprotein-protein interactions, including interactions between lin-8,lin-56, and lin-61 gene products and other synMuv gene products. Forexample, in a genome-wide yeast two-hybrid screen, using LIN-35 as abait, we showed LIN-8 and LIN-35 to interact.

[0142] Interactions among the lin-8, lin-56, lin-61 and other synMuvgene products can also be examined using other methods known to oneskilled in the art. For example such interactions can be determined byperforming GST pull-down experiments using the appropriate fusionproteins, as described in U.S. Ser. No. 09/220,091. Alternatively,interactions may be further investigated through the use of triplemutants using the appropriate genes, as also described in U.S. Ser. No.09/220,091.

[0143] 6. Non-Vulval Phenotypes in lin-8 and lin-61 Mutants

[0144] lin-8 and lin-6 are also involved in a non-vulval phenotype.During the course of a C. elegans screen using a cell-type specificreporter, which results in the expression of green fluorescent protein,we discovered that a class of mutants exists in which this reporter isectopically expressed in pharyngeal tissue, especially in the posteriorpharynx. We refer to this phenotype to as the “green pharynx phenotype.”Further studies have revealed that loss-of-function mutations in any ofseveral genes (lin-8, lin-13, lin-61) belonging to both the class A andclass B synMuv pathways can cause the green pharynx phenotype. In thecase of lin-8, all but one allele (n2376, E148K) tested exhibit thegreen pharynx phenotype, providing a single, distinct loss-of-functionphenotype for this gene.

[0145] In addition, some of the synMuv B genes are homologues of NuRDcomplex components and loss-of-function mutations in other components ofthe NuRD chromatin remodeling complex (e.g., lin-40/egr-1 and chd-3) canalso cause the green pharynx phenotype. Furthermore, the green pharynxphenotype has been observed with two distinct cell fate-specificreporters (pkd-2::gfp and lin-11::gfp).

[0146] The green pharynx phenotype is observed irrespective of whetherthe reporter transgene is integrated into the genome or whether it ispresent extra-chromosomally, indicating that chromosomal integration isnot required. Additionally, when using an integrated reporter, thephenotype does not appear to be highly dependent on the site ofchromosomal integration and the phenotype is observed with both low andhigh copy number transgenes.

[0147] Our data indicate that the synMuv A and B genes may act insimilar processes, and may work together in contexts other than vulvaldevelopment and provide additional evidence that lin-8, lin-13 andlin-61 act in transcriptional repression.

[0148] 7. Cloning lin-8, lin-56 and lin-61 Vertebrate Genes

[0149] The invention described herein provides the identity of class AsynMuv genes lin-8 and lin-56, and the class B synMuv gene lin-61. Inview of what is know in the field regarding the synMuv family and itsrelationship to related genes in the Rb tumor suppressor pathway, weconclude that these newly cloned genes are also involved in pathwaysthat modulate tumor suppression. It is likely that C. elegans lin-8,lin-56 and lin-61 genes have ortholog counterparts in vertebrates, forexample mammalian genes. One skilled in the art will recognize thatthese orthologs can be identified using standard techniques in molecularbiology, such as screening of cDNA or genomic libraries, degenerate PCR,and the like, described in, for example, Ausubel et al. (supra).Orthologs can also be identified using computer-based search programssuch as BLAST and dbEST to isolate expressed sequence tags (ESTs) withregions of similarity or identity to a lin-8, lin-56 or lin-61 gene.

[0150] Vertebrate counterparts of C elegans lin-8, lin-56 and lin-61 arecandidate tumor suppressor genes. Thus, one can screen for mutations inthe human homologs of these genes in patients diagnosed with cancer orin immortalized cell lines. Similarly, the polypeptides encoded by thesegenes are candidate targets for anti-cancer drugs. A drug whichincreases synMuv polypeptide activity, for example, LIN-8, LIN-56, orLIN-61 biological activity, may decrease proliferation of tumor cells.In addition, polypeptides which interact with other synMuv polypeptidesor which regulate synMuv gene expression are also candidate tumorsuppressors; these polypeptides can be isolated using standardtechniques, as described herein or, for example, in Ausubel et al.(supra).

[0151] 8. LIN-8, LIN-56, or LIN-61 Polypeptide Expression

[0152] A lin-8, lin-56, or lin-61 nucleic acid sequence may be expressedin a prokaryotic or eukaryotic cell. In addition, it may be desirable toexpress the nucleic acid sequence under the control of an induciblepromoter for the purposes of polypeptide production.

[0153] In general, LIN-8, LIN-56, or LIN-61 polypeptides may be producedby transformation of a suitable host cell with all or part of a LIN-8,LIN-56, or LIN-61-encoding cDNA fragment (e.g., the cDNAs describedabove) in a suitable expression vehicle.

[0154] Those skilled in the field of molecular biology will understandthat any of a wide variety of expression systems may be used to providethe recombinant protein. The precise host cell used is not critical tothe invention. The LIN-8, LIN-56, or LIN-61 polypeptide may be producedin a prokaryotic host (e.g., E. coli) or in a eukaryotic host (e.g.,nematodes, Saccharomyces cerevisiae, insect cells, e.g., Sf-21 cells, ormammalian cells, e.g., COS 1, NIH 3T3, or HeLa cells). Such cells areavailable from a wide range of sources (e.g., the American Type CultureCollection, Rockland, Md.; also, see, e.g., Ausubel et al. (supra)). Themethod of transformation or transfection and the choice of expressionvehicle will depend on the host system selected. Transformation andtransfection methods are described, e.g., in Ausubel et al. (supra);expression vehicles may be chosen from those provided, e.g., in CloningVectors: A Laboratory Manual (P. H. Pouwels et al., 1985, Supp. 1987).

[0155] One preferred expression system is the baculovirus system (using,for example, the vector pBacPAK9) available from Clontech (Palo Alto,Calif.). If desired, this system may be used in conjunction with otherprotein expression techniques, for example, the myc tag approachdescribed by Evan et al. (Mol. Cell Biol. 5:3610-3616, 1985).

[0156] Alternatively, a LIN-8, LIN-56, or LIN-61 polypeptide is producedby a stably-transfected mammalian cell line. A number of vectorssuitable for stable transfection of mammalian cells are available to thepublic, e.g., see Pouwels et al. (supra); methods for constructing suchcell lines are also publicly available, e.g., in Ausubel et al. (supra).In one example, cDNA encoding a LIN-8, LIN-56, or LIN-61 polypeptide iscloned into an expression vector which includes the dihydrofolatereductase (DHFR) gene. Integration of the plasmid, and, therefore, theLIN-8, LIN-56, or LIN-61 polypeptide-encoding gene, into the host cellchromosome is selected for by inclusion of 0.01-300 μM methotrexate inthe cell culture medium (as described in Ausubel et al. (supra)). Thisdominant selection can be accomplished in most cell types andrecombinant protein expression can be increased by DHFR-mediatedamplification of the transfected gene. In addition, methods forselecting cell lines bearing gene amplifications are described inAusubel et al. (supra); such methods generally involve extended culturein medium containing gradually increasing levels of methotrexate.DHFR-containing expression vectors commonly used for this purposeinclude pCVSEII-DHFR and pAdD26SV(A) (described in Ausubel et al.(supra)). Any of the host cells described above or, preferably, aDHFR-deficient CHO cell line (e.g., CHO DHFR⁽⁻⁾ cells, ATCC AccessionNo. CRL 9096) are among the host cells preferred for DHFR selection of astably-transfected cell line or DHFR-mediated gene amplification.

[0157] Once the recombinant LIN-8, LIN-56, or LIN-61 protein isexpressed, it is isolated, e.g., using affinity chromatography. In oneexample, an anti-LIN-8, LIN-56, or LIN-61 protein antibody (e.g.,produced as described herein) may be immobilized on a column and used toisolate the LIN-8, LIN-56, or LIN-61 protein. Lysis and fractionation ofLIN-8, LIN-56, or LIN-61 protein-harboring cells prior to affinitychromatography may be performed by standard methods (see, e.g., Ausubelet al. (supra)).

[0158] Once isolated, the recombinant protein can, if desired, befurther purified, e.g., by high pressure liquid chromatography (see,e.g., Fisher, Laboratory Techniques In Biochemistry And MolecularBiology, eds., Work and Burdon, Elsevier, 1980).

[0159] Polypeptides of the invention, particularly short LIN-8, LIN-56,or LIN-61 protein fragments, can also be produced by chemical synthesis(e.g., by the methods described in Solid Phase Peptide Synthesis, 2nded., 1984, The Pierce Chemical Co., Rockford, Ill.).

[0160] These general techniques of polypeptide expression andpurification can also be used to produce and isolate useful LIN-8,LIN-56, or LIN-61 fragments or analogs (described herein).

[0161] 9. Anti-LIN-8, LIN-56, or LIN-61 Antibodies

[0162] In general, to generate a LIN-8, LIN-56, or LIN-61-specificantibody, a lin-8, lin-56, or lin-61 coding sequence may be expressed asa C-terminal fusion with glutathione S-transferase (GST) (Smith et al.,Gene 67:31-40, 1988). The fusion protein can be purified onglutathione-sepharose beads, eluted with glutathione cleaved withthrombin (at the engineered cleavage site), and purified to the degreenecessary for immunization of rabbits. Primary immunizations can becarried out with Freund's complete adjuvant and subsequent immunizationswith Freund's incomplete adjuvant. Antibody titres are monitored byWestern blot and immunoprecipitation analyses using the thrombin-cleavedLIN-8, LIN-56, or LIN-61 polypeptide fragment of the GST-LIN-8, -LIN-56,or -LIN-61 fusion protein. Immune sera are affinity purified using, forexample, CNBr-Sepharose-coupled LIN-8, LIN-56, or LIN-61 protein.Antiserum specificity is determined using a panel of unrelated GSTproteins (including GSTp53, Rb, HPV-16 E6, and E6-AP) and GST-trypsin(which was generated by PCR using known sequences).

[0163] As an alternate or adjunct immunogen to GST fusion proteins,peptides corresponding to relatively unique regions of LIN-8, LIN-56, orLIN-61 may be generated and coupled to keyhole limpet hemocyanin (KLH)through an introduced C-terminal lysine. Antiserum to each of thesepeptides is similarly affinity purified on peptides conjugated to BSA,and specificity tested in ELISA and Western blots assays using peptideconjugates, and by Western blot and immunoprecipitation techniques usingLIN-8, LIN-56, or LIN-61 expressed as a GST fusion protein.

[0164] Alternatively, monoclonal antibodies may be prepared using theLIN-8, LIN-56, or LIN-61 proteins described above and standard hybridomatechnology (see, e.g., Kohler et al., Nature 256:495, 1975; Kohler etal., Eur. J. Immunol. 6:511, 1976; Kohler et al., Eur. J. Immunol.6:292, 1976; Hammerling et al., In Monoclonal Antibodies and T CellHybridomas, Elsevier, N.Y., 1981; Ausubel et al., (supra)). Onceproduced, monoclonal antibodies are also tested for specific recognitionby Western blot or immunoprecipitation analysis (by the methodsdescribed in Ausubel et al. (supra). Antibodies which specificallyrecognize LIN-8, LIN-56, or LIN-61 are considered to be useful in theinvention; such antibodies may be used, e.g., in an immunoassay tomonitor the level of LIN-8, LIN-56, or LIN-61 produced by an animal (forexample, to determine the amount or subcellular location of LIN-8,LIN-56, or LIN-61).

[0165] Preferably antibodies of the invention are produced usingfragments of the LIN-8, LIN-56, or LIN-61 polypeptide that arepositioned outside highly conserved regions and appear likely to beantigenic, by criteria such as those provided by the Peptide structureprogram of the Genetics Computer Group Sequence Analysis Package(Program Manual for the GCG Package, Version 7, 1991) using thealgorithm of Jameson and Wolf (CABIOS 4:181, 1988). In one specificexample, such fragments are generated by standard PCR techniques andcloned into the pGEX expression vector (Ausubel et al. (supra). Fusionproteins are expressed in E. coli and purified using a glutathioneagarose affinity matrix, as described in Ausubel et al. (supra). Toattempt to minimize the potential problems of low affinity orspecificity of antisera, two or three such fusions are generated foreach polypeptide, and each fusion is injected into at least two rabbits.Antisera are raised by injections in a series, preferably including atleast three booster injections.

[0166] To demonstrate the utility of this approach, we generatedpolyclonal antibodies against a fusion of full-length LIN-8 with maltosebinding protein (MBP) (see Example 1 for the detailed protocol used). Inorder to increase the sensitivity of the antibodies, they wereaffinity-purified against a GST::LIN-8 fusion, and pre-adsorbed withextract from lin-8(n2731) worms.

[0167] In addition, the antibodies that we generated against LIN-56 (seeExample 2) were also affinity purified and pre-adsorbed with extractfrom lin-56(n2728) worms. We used one of these anti-LIN-56 antibodies(HM1923) for Western analysis and for wholemount staining. This antibodyrecognizes a doublet in wild-type but not lin-56(n2728) worm extracts onWestern analysis and the proteins in this doublet fractionatespecifically with nuclear material. Furthermore, wholemount stainingwith this antibody reveals that lin-56 is expressed in the nuclei ofmost if not all cells throughout development and adulthood (FIG. 9 A-D).We also stained lin-56(n2728) embryos, larvae, and adults to show thatthe HM1923 antibody is specific for LIN-56. As is seen in FIG. 10,LIN-56 staining is absent in LIN-56 mutant worms, indicating that theantibody is specific for LIN-56.

[0168] In addition, LIN-56 appears to be absent from nuclei during partof the cell cycle, probably as a result of nuclear membrane breakdown.Furthermore, LIN-56 expression and localization appear wild-type in thesynMuv A lin-8 and lin-38 mutants, but the nuclear expression of LIN-56appears severely reduced or even absent in lin-1SA(n767) andlin-15AB(n309) mutants. By Western analysis, LIN-56 protein levels do infact appear reduced in lin-15A(n 767) worm extracts, and the ratio ofthe bands in the detected doublet may even be altered.

[0169] 10. Identification of Molecules that Modulate LIN-8, LIN-56, orLIN-61 Polypeptide Expression

[0170] Isolation of lin-8, lin-56, or lin-61 cDNAs also facilitates theidentification of molecules which increase or decrease LIN-8, LIN-56, orLIN-61 expression. According to one approach, candidate molecules areadded at varying concentrations to the culture medium of cells ornematodes expressing LIN-8, LIN-56, or LIN-61. lin-8, lin-56, or lin-61expression is then measured, for example, by standard Northern blotanalysis (Ausubel et al. (supra)) using a lin-8, lin-56, or lin-61 cDNA(or cDNA fragment) as a hybridization probe (see also Table III). Thelevel of lin-8, lin-56, or lin-61 expression in the presence of thecandidate molecule is compared to the level measured for the same cellsin the same culture medium but in the absence of the candidate molecule.When nematodes are being used, the phenotypes associated with the synMuvpathway may be utilized as the primary screen for alteration inpolypeptide expression.

[0171] If desired, the effect of candidate modulators on expression may,in the alternative, be measured at the level of LIN-8, LIN-56, or LIN-61polypeptide production using the same general approach and standardimmunological detection techniques, such as Western blotting orimmunoprecipitation with a LIN-8, LIN-56, or LIN-61-specific antibody(for example, the LIN-8, LIN-56, or LIN-61 antibody described herein).

[0172] Candidate modulators may be purified (or substantially purified)molecules or may be one component of a mixture of compounds (e.g., anextract or supernatant obtained from cells; Ausubel et al. (supra)). Ina mixed compound assay, LIN-8, LIN-56, or LIN-61 expression is testedagainst progressively smaller, subsets of the candidate compound pool(e.g., produced by standard purification techniques, e.g., HPLC or FPLC)until a single compound or minimal compound mixture is demonstrated tomodulate LIN-8, LIN-56, or LIN-61 expression.

[0173] Alternatively, or in addition, candidate compounds may bescreened for those, which modulate LIN-8, LIN-56, or LIN-61 cellproliferation. In this approach, the degree of cell proliferation, orthe LIN-8, LIN-56, or LIN-61 phenotype in the presence of a candidatecompound is compared to the degree of cell proliferation in its absence,under equivalent conditions. Again, such a screen may begin with a poolof candidate compounds, from which one or more useful modulatorcompounds are isolated in a step-wise fashion. Cell proliferation may bemeasured by any standard assay.

[0174] Candidate LIN-8, LIN-56, or LIN-61 modulators include peptide aswell as non-peptide molecules (e.g., peptide or non-peptide moleculesfound, e.g., in a cell extract, mammalian serum, or growth medium onwhich mammalian cells have been cultured).

[0175] Modulators found to be effective at the level of LIN-8, LIN-56,or LIN-61 expression or biological activity may be confirmed as usefulin animal models and, if successful, may be used as anti-cancertherapeutics to increase or decrease cell proliferation.

[0176] 11. lin-8, lin-56, or lin-61 Therapy

[0177] Because expression levels of lin-8, lin-56, or lin-61 genescorrelate with the levels of cell proliferation, such genes also finduse in gene therapy to modulate cell proliferation.

[0178] Retroviral vectors, adenoviral vectors, adeno-associated viralvectors, or other viral vectors with the appropriate tropism for cellslikely to be involved in the cell proliferation disease may be used as agene transfer delivery system for a therapeutic lin-8, lin-56, or lin-61gene construct. Numerous vectors useful for this purpose are generallyknown in the art (Miller, Human Gene Therapy 5-14, 1990; Friedman,Science 244:1275-1281, 1989; Eglitis and Anderson, BioTechniques6:608-614, 1988; Tolstoshev and Anderson, Current Opinion inBiotechnology 1:55-61, 1990; Sharp, The Lancet 337:1277-1278, 1991;Cornetta et al., Nucleic Acid Research and Molecular Biology 36:311-322,1987; Anderson, Science 226:401-409, 1984; Moen, Blood Cells 17:407-416,1991; and Miller and Rosman, BioTechniques 7:980-990, 1989; Le Gal LaSalle et al., Science 259:988-990, 1993; and Johnson, Chest 107:77S-83S,1995). Retroviral vectors are particularly well developed and have beenused in clinical settings (Rosenberg et al., N. Engl. J. Med 323:370,1990; Anderson et al., U.S. Pat. No. 5,399,346).

[0179] Non-viral approaches may also be employed for the introduction oftherapeutic DNA into cells otherwise predicted to undergo insufficientor excess cell proliferation. For example, lin-8, lin-56, or lin-61 maybe introduced into a cell by the techniques of lipofection (Felgner etal., Proc. Natl. Acad. Sci. USA 84:7413, 1987; Ono et al., NeuroscienceLett. 117:259, 1990; Brigham et al., Am. J. Med. Sci. 298:278, 1989;Staubinger and Papahadjopoulos, Meth. Enz. 101:512, 1983);asialorosonucoid-polylysine conjugation (Wu and Wu, J. Biol. Chem.263:14621, 1988; Wu et al., J. Biol. Chem. 264:16985, 1989); or, lesspreferably, microinjection under surgical conditions (Wolff et al.,Science 247:1465, 1990).

[0180] For any of the above approaches, the therapeutic lin-8, lin-56,or lin-61 DNA construct, or an antisense nucleic acid, is preferablyapplied to the site of the predicted cell proliferation event (forexample, by injection), but may also be applied to tissue in thevicinity of the predicted event or even to a blood vessel supplying thecells predicted to undergo insufficient or excess cell proliferation.

[0181] In the gene therapy constructs, lin-8, lin-56, or lin-61 cDNAexpression is directed from any suitable promoter (e.g., the humancytomegalovirus, simian virus 40, or metallothionein promoters), and itsproduction is regulated by any desired regulatory element. For example,if desired, enhancers known to direct preferential gene expression in aparticular cell may be used to direct lin-8, lin-56, or lin-61expression. Such enhancers include, without limitation, those enhancerswhich are characterized as tissue or cell specific in their expression.

[0182] Alternatively, if a lin-8, lin-56, or lin-61 genomic clone isutilized as a therapeutic construct (for example, following itsisolation by hybridization with the lin-8, lin-56, or lin-61 cDNAdescribed above), lin-8, lin-56, or lin-61 expression is regulated byits cognate regulatory sequences or, if desired, by regulatory sequencesderived from a heterologous source, e.g., any of the promoters orregulatory elements described above.

[0183] Less preferably, lin-8, lin-56, or lin-61 gene therapy isaccomplished by direct administration of the lin-8, lin-56, or lin-61mRNA to a cell predicted to undergo excess or insufficient cellproliferation. This mRNA may be produced and isolated by any standardtechnique, but is most readily produced by in vitro transcription usinga lin-8, lin-56, or lin-61 cDNA under the control of a high efficiencypromoter (e.g., the T7 promoter). Administration of lin-8, lin-56, orlin-61 mRNA to malignant cells is carried out by any of the methods fordirect nucleic acid administration described above.

[0184] Ideally, the production of a LIN-8, LIN-56, or LIN-61 polypeptideby any gene therapy approach described above results in a cellular levelof LIN-8, LIN-56, or LIN-61 that is at least equivalent to the normal,cellular level of LIN-8, LIN-56, or LIN-61 in an unaffected individual.Treatment by any lin-8, lin-56, or lin-61-mediated gene therapy approachmay be combined with more traditional therapies.

[0185] Another therapeutic approach included within the inventioninvolves direct administration of recombinant LIN-8, LIN-56, or LIN-61protein, either to the site of a predicted or desirable cellproliferation event (for example, by injection) or systemically by anyconventional recombinant protein administration technique. The actualdosage of LIN-8, LIN-56, or LIN-61 administered depends on a number offactors, including the size and health of the individual patient, but,generally, between 0.1 mg and 100 mg, inclusive, are administered perday to an adult in any pharmaceutically-acceptable formulation.

[0186] The nucleic acids of the present invention may also be utilizedin plant cells. Such sequences may be expressed in plant cells, andused, for example, to promote plant survival or growth (e.g., byproviding disease resistance).

[0187] 12. Administration of LIN-8 LIN-56, or LIN-61 Polypeptides,lin-8, lin-56, or lin-61 Nucleic Acid Sequences, or Modulators of LIN-8,LIN-56, or LIN-61 Synthesis or Function

[0188] A LIN-8, LIN-56, or LIN-61 polypeptide, nucleic acid sequence, ormodulator may be administered with a pharmaceutically-acceptablediluent, carrier, or excipient, in unit dosage form. Conventionalpharmaceutical practice may be employed to provide suitable formulationsor compositions to administer LIN-8, LIN-56, or LIN-61 to patientssuffering from, or presymptomatic for, a LIN-8, LIN-56, or LIN-61, orsynMuv-associated cancer. Any appropriate route of administration may beemployed, for example, parenteral, intravenous, subcutaneous,intramuscular, intracranial, intraorbital, ophthalmic, intraventricular,intracapsular, intraspinal, intracistemal, intraperitoneal, intranasal,aerosol, or oral administration. Therapeutic formulations may be in theform of liquid solutions or suspensions; for oral administration,formulations may be in the form of tablets or capsules; and forintranasal formulations, in the form of powders, nasal drops, oraerosols.

[0189] Methods well known in the art for making formulations are found,for example, in Remington's Pharmaceutical Sciences ((18^(th) edition),ed. A. Gennaro, 1990, Mack Publishing Company, Easton, Pa.).Formulations for parenteral administration may, for example, containexcipients, sterile water, or saline, polyalkylene glycols such aspolyethylene glycol, oils of vegetable origin, or hydrogenatednapthalenes. Biocompatible, biodegradable lactide polymer,lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylenecopolymers may be used to control the release of the compounds. Otherpotentially useful parenteral delivery systems for LIN-8, LIN-56, orLIN-61 polypeptides, nucleic acid sequences or modulatory compoundsinclude ethylene-vinyl acetate copolymer particles, osmotic pumps,implantable infusion systems, and liposomes. Formulations for inhalationmay contain excipients, for example, lactose, or may be aqueoussolutions containing, for example, polyoxyethylene-9-lauryl ether,glycocholate and deoxycholate, or may be oily solutions foradministration in the form of nasal drops, or as a gel.

[0190] If desired, treatment with a LIN-8, LIN-56, or LIN-61polypeptide, nucleic acid sequence, or modulatory compound may becombined with more traditional therapies for the disease such assurgery, radiation, or chemotherapy for cancers.

[0191] 13. Detection of A Condition Involving Altered Cell Proliferationor an Increased Likelihood of Developing a Cell Proliferation Disease

[0192] LIN-8, LIN-56, or LIN-61 polypeptides and nucleic acid sequencesfind diagnostic use in the detection or monitoring of conditionsinvolving aberrant levels of cell proliferation. A decrease or increasein the level of LIN-8, LIN-56, or LIN-61 production may provide anindication of a deleterious condition. Levels of LIN-8, LIN-56, orLIN-61 expression may be assayed by any standard technique. For example,its expression in a biological sample (e.g., a biopsy) may be monitoredby standard Northern blot analysis, using, for example, probes designedfrom lin-8, lin-56, or lin-61 nucleic acid sequences, or from nucleicacid sequences that hybridize to a lin-8, lin-56, or lin-61 nucleic acidsequence. Measurement of such expression may be aided by PCR (see, e.g.,Ausubel et al. (supra); PCR Technology: Principles and Applications forDNA Amplification, ed., H. A. Ehrlich, Stockton Press, NY; and Yap andMcGee, Nucl. Acids Res. 19:4294, 1991).

[0193] Alternatively, a patient sample may be analyzed for one or moremutations in the lin-8, lin-56, or lin-61 sequences using a mismatchdetection approach. Generally, these techniques involve PCRamplification of nucleic acid from the patient sample, followed byidentification of the mutation (i.e., mismatch) by either alteredhybridization, aberrant electrophoretic gel migration, binding orcleavage mediated by mismatch binding proteins, or direct nucleic acidsequencing. Any of these techniques may be used to facilitate mutantlin-8, lin-56, or lin-61 detection, and each is well known in the art(see, for example, Orita et al., Proc. Natl. Acad. Sci. USA86:2766-2770, 1989; and Sheffield et al., Proc. Natl. Acad. Sci. USA86:232-236, 1989).

[0194] In yet another approach, immunoassays are used to detect ormonitor a LIN-8, LIN-56, or LIN-61 polypeptide in a biological sample.LIN-8, LIN-56, or LIN-61-specific polyclonal or monoclonal antibodies(produced as described above) may be used in any standard immunoassayformat (e.g., ELISA, Western blot, or RIA assay) to measure LIN-8,LIN-56, or LIN-61 polypeptide levels; again comparison is to wild-typeLIN-8, LIN-56, or LIN-61 levels, and an increase or decrease in LIN-8,LIN-56, or LIN-61 production is indicative of a condition involvingaltered cell proliferation. Examples of immunoassays are described,e.g., in Ausubel et al. (supra). Immunohistochemical techniques may alsobe utilized for LIN-8, LIN-56, or LIN-61 detection. For example, atissue sample may be obtained from a patient, and a section stained forthe presence of LIN-8, LIN-56, or LIN-61 using an anti-LIN-8, LIN-56, orLIN-61 antibody and any standard detection system (e.g., one whichincludes a secondary antibody conjugated to horseradish peroxidase).General guidance regarding such techniques can be found in, e.g.,Bancroft and Stevens (Theory and Practice of Histological Techniques,Churchill Livingstone, 1982) and Ausubel et al. (supra).

[0195] In one preferred example, a combined diagnostic method may beemployed that begins with an evaluation of LIN-8, LIN-56, or LIN-61polypeptide production (for example, by immunological techniques or theprotein truncation test (Hogerrorst et al., Nature Genetics 10:208-212,1995) and also includes a nucleic acid-based detection techniquedesigned to identify more subtle lin-8, lin-56, or lin-61 mutations (forexample, point mutations). As described above, a number of mismatchdetection assays are available to those skilled in the art, and anypreferred technique may be used (see above). By this approach, mutationsin lin-8, lin-56, or lin-61 may be detected that either result in lossof LIN-8, LIN-56, or LIN-61 expression or biological activity.

[0196] Mismatch detection assays also provide the opportunity todiagnose a lin-8, lin-56, or lin-61-mediated predisposition to diseasesof cell proliferation. For example, a patient heterozygous for a lin-8,lin-56, or lin-61 mutation may show no clinical symptoms and yet possessa higher than normal probability of developing one or more types ofdiseases. Given this diagnosis, a patient may take precautions tominimize their exposure to adverse environmental factors (for example,UV exposure or chemical mutagens) and to carefully monitor their medicalcondition (for example, through frequent physical examinations). Thistype of lin-8, lin-56, or lin-61 diagnostic approach may also be used todetect lin-8, lin-56, or lin-61 mutations in prenatal screens.

[0197] The lin-8, lin-56, or lin-61 diagnostic assays described abovemay be carried out using any biological sample (for example, any biopsysample or bodily fluid or tissue) in which lin-8, lin-56, or lin-61 isnormally expressed. Identification of a mutant lin-8, lin-56, or lin-61gene may also be assayed using these sources for test samples.Alternatively, a lin-8, lin-56, or lin-61 mutation, particularly as partof a diagnosis for predisposition to lin-8, lin-56, or lin-61-associatedproliferative disease, may be tested using a DNA sample from any cell,for example, by mismatch detection techniques; preferably, the DNAsample is subjected to PCR amplification prior to analysis.

[0198] The following examples are meant to illustrate the invention andshould not be construed as limiting.

EXAMPLES Example 1 Generation of Rabbit and Guinea Pig PolyclonalAntibodies Against LIN-8

[0199] We made a fusion protein of full-length LIN-8 with MBP and hadCovance (Richmond, Calif.) produce LIN-8 polyclonal antibodies using tworabbits and two guinea pigs. However, anyone skilled in the art maygenerate antibodies against LIN-8 using the following protocol.

[0200] New Zealand White Female Rabbits were bled prior to injection ofthe antigen, for later use as a control in establishing backgroundreactivity of the serum. Following the prebleed, the rabbits wereinjected subcutaneously at multiple sites with 250 μg protein and 0.5 mLFreund's Complete Adjuvant (FCA). Three weeks later, the rabbitsreceived a subcutaneous, dorsal, boost injection of 125 μg protein with1.0 mL Freund's Incomplete Adjuvant (FIA). The first test bleed wasperformed 11 days later, followed by a second subcutaneous, dorsal boostinjection of 125 μg protein and 1.0 mL FIA 9 days after the test bleed.A second test bleed was performed 11 days after the second boost,followed by a third subcutaneous, dorsal boost of 125 μg protein and 1.0mL FIA 10 days after the second test bleed. The first production bleedwas performed 10 days after the third boost, followed by a fourthsubcutaneous, dorsal boost of 125 μg and 1.0 mL FIA 10 days after thefirst production bleed. A second production bleed was performed 11 daysafter the fourth boost, followed by a fifth subcutaneous, dorsal boostof 125 μg and 1.0 mL FIA 10 days after the second production bleed. Athird production bleed was performed 11 days after the fifth boost,followed by exsanguination after 6 days.

[0201] The protocol used to produce LIN-8 antibodies in guinea pigsclosely follows the one outlined for rabbits above. Dunkin HartleyGuinea Pig were used and prebled prior to subcutaneous and intradermalinjection of 200 μg protein and 1.0 ml FCA. Three weeks after theprimary injection, the guinea pigs received a boost injection of 100 μgprotein and 0.5 mL FIA, subcutaneously in the neck. After 11 days, thefirst test bleed was performed, followed 10 days later by a second boostof 100 μg protein and 0.5 mL FIA, subcutaneously in the neck. The secondtest bleed was performed after 11 days, followed 10 days later by athird boost of 100 μg protein and 0.5 mL FIA, subcutaneously in theneck. The first production bleed was performed 11 days after the thirdboost, followed 10 days later by a fourth subcutaneous, dorsal boost of100 μg protein and 0.5 mL FIA. The second production bleed followed 11days later and a fifth boost of 100 μg protein and 0.5 mL FIA wasperformed after 10 days. The third production bleed was performed 11days later, followed by exsanguination after 6 days.

[0202] Generally, polyclonal antibodies are affinity purified toincrease their specificity. In addition, the polyclonal antibody may bedepleted of any components that do not specifically bind to the proteinof interest by pre-adsorbing the antibody with an extract made fromtissue that lacks the protein of interest. For example, an extract fromlin-8(n2731) worms may be used to remove any antibodies that bind wormproteins besides LIN-8.

Example 2 Generation of Rabbit and Rat Polyclonal Antibodies AgainstLIN-56

[0203] We made a fusion protein of full-length LIN-56 with GST and hadCovance (Richmond, Calif.) produce LIN-56 polyclonal antibodies usingtwo rabbits and two rats. However, anyone skilled in the art maygenerate antibodies against LIN-56 using the following protocol.

[0204] New Zealand White Female Rabbits were bled prior to intradermalinjection in the back with 250 μg protein and 0.5 mL FCA. Three weekslater, the rabbits received a subcutaneous nodal (groin and pit) areaboost injection of 125 μg protein with 0.5 mL FIA. The first test bleedwas performed 10 days later, followed by a second subcutaneous boostinjection of 125 μg protein and 0.5 mL FIA in the neck, 11 days afterthe test bleed. A second test bleed was performed 10 days after thesecond boost, followed by a third subcutaneous, dorsal boost of 125 μgprotein and 1.0 mL FIA 11 days after the second test bleed. The firstproduction bleed was performed 10 days after the third boost, followedby a fourth subcutaneous nodal area (groin and pit) boost of 125 μg and1.0 mL FIA 10 days after the first production bleed. A second productionbleed was performed 11 days after the fourth boost, followed by a fifthsubcutaneous, dorsal boost of 125 μg and 1.0 mL FIA 11 days after thesecond production bleed. A third production bleed was performed 10 daysafter the fifth boost, followed by exsanguination after 10 days.

[0205] The protocol used to produce LIN-56 antibodies in rats closelyfollows the one outlined for rabbits above. SD rats were used andprebled prior to subcutaneous injection of 200 μg protein and 0.4 mLFCA, in the neck. Three weeks after the primary injection, the ratsreceived a boost injection of 100 μg protein and 0.4 mL FIA,subcutaneously in the neck. After 10 days, the first test bleed wasperformed, followed 11 days later by a second boost of 100 μg proteinand 0.4 mL FIA, subcutaneously in the neck. The second test bleed wasperformed after 10 days, followed 11 days later by a third boost of 100μg protein and 0.4 mL FIA, subcutaneously in the neck. The firstproduction bleed was performed 10 days after the third boost, followed11 days later by a fourth subcutaneous, dorsal boost of 100 μg proteinand 0.4 mL FIA. The second production bleed followed 10 days later and afifth boost of 100 μg protein and 0.4 mL FIA was performed after 11days. The third production bleed was performed 10 days later, followedby exsanguination after 10 days.

Other Embodiments

[0206] While the invention has been described in connection withspecific embodiments thereof, it will be understood that it is capableof further modifications. This application is intended to cover anyvariations, uses, or adaptations of the invention following, in general,the principles of the invention and including such departures from thepresent disclosure as come within known or customary practice within theart to which the invention pertains and which may be applied to theessential features hereinbefore set forth.

[0207] All publications and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each independent publication or patent application was specificallyand individually indicated to be incorporated by reference.

1 78 1 386 PRT Caenorhabditis elegans 1 Met Ser Lys Ile Lys Thr His SerThr Gly Ser Lys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro ProVal Pro Leu Pro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe SerThr Glu Lys Tyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe AspAsp Tyr Asp Val Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn GluIle Gly Lys Cys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala AlaIle Met Glu His Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg ThrGly Leu Leu Leu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys CysGly Lys Asp Asn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val SerLys Arg Leu Thr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg TrpGlu Phe Tyr Gly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 GlnArg Trp Glu Ala Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175Leu Glu Ala Arg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly 180 185190 Glu Leu Met Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp Glu Met 195200 205 Cys Val Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn Trp Ser Asp210 215 220 Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser Pro Glu AlaLys 225 230 235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala Asp Glu ValTyr Asn Pro 245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu Ser Ala SerAsn Ala Met Tyr 260 265 270 Arg Ile Ala Phe Ser Gln Gln Tyr Gly Gly GlyGly Ser Pro Ala Val 275 280 285 Gln Lys Pro Val Thr Phe Ser Ala Gln ProAla Pro Ala Pro Val Arg 290 295 300 Glu Ala Pro Ser Pro Val Val Glu AsnVal Ser Ser Ser Ser Phe Thr 305 310 315 320 Pro Lys Pro Pro Ala Met IleAsn Asn Phe Gly Glu Glu Met Asn Gln 325 330 335 Ile Thr Tyr Gln Ala IleArg Ile Ala Arg Glu Gln Pro Glu Arg Leu 340 345 350 Lys Leu Leu Arg LysAla Leu Phe Asp Val Val Leu Ala Phe Asp Gln 355 360 365 Lys Glu Tyr AlaAsp Val Gly Asp Leu Tyr Arg Asp Leu Ala Gln Lys 370 375 380 Asn Ser 3852 1276 DNA Caenorhabditis elegans 2 agaatctgcc aaaatgtcaa agataaagacacattccact ggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccacttccaccactc ccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcactgagcaaagat gagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaaagtggtgctc aacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagccattatggag cactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgttgtctatcaaa tcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggcttcgcgtggca attgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcgctgggagttt tacggcttta ttcgctacta 480 tcgagactat acacaacgct gggaggccgacttgttgaaa gatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatatggaaaaggtg gattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatggatgagatgtgc gtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggcgccggaacca tcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacctactacctgag gcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatccgcatcaaac gccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtccccagccgtgcag aagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggccccaagccca gttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccatgatcaacaat tttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccgagagcagccg gaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtttgatcagaag gaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattcgtgataattt ttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttccaatataata aacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 3 322 PRTCaenorhabditis elegans 3 Met Asp His His Ala Met Tyr Arg Thr Ala Glu PheAsn Lys Thr Thr 1 5 10 15 Val Arg Leu Leu Ala Glu Phe Ile Glu Lys ThrGly Gln Asn Ala Thr 20 25 30 Ile Val Asn Met Asp Ser Phe Leu Glu Phe PheAla Tyr Leu Asn Pro 35 40 45 Thr Ala Pro Ile Pro Thr Val Pro Glu Ile GluLys Gln Leu Leu Leu 50 55 60 Lys Ser Pro Ile Arg Cys Ile Val Cys Gly MetGlu Thr Glu Ser Asp 65 70 75 80 Ser Ala Val Thr Leu Ser Ile Asp Asn AlaSer Ile Ile Leu Thr Ala 85 90 95 Thr Val Ile Gly Tyr Cys Arg Asp Pro SerAsp Ala Val Asn Gln Ile 100 105 110 Arg Lys Glu Ser Leu Arg Ala Cys ThrLys His Phe Asn Ser Ile Phe 115 120 125 His Val Ile Phe Glu Gly Leu GlnIle Glu Asn Thr Tyr Cys Ala His 130 135 140 His Ala Lys Tyr Ser Leu AlaAsn Arg Trp Cys Lys Val Tyr Thr Met 145 150 155 160 Ile Arg Ser Ser LeuGly Glu Gln Phe Thr Lys Phe Asp Val Arg Asn 165 170 175 Phe Lys Ser IleLeu Gln Ser Phe Leu Asp Thr Phe Gly Glu Ile Asp 180 185 190 Asp Asp LysLys Asp Lys Glu Ser Ser His Phe Asp Glu Cys Phe Glu 195 200 205 Glu MetAsp Ser Glu Asn Val Glu Ile Lys Met Glu Ser Pro Gln Glu 210 215 220 GluAla Ala Glu Lys Ser Lys Phe Ser Glu Asn Leu Val Glu Val Lys 225 230 235240 Leu Glu Pro Ile Glu Thr His Glu Leu Asp Lys Thr Ile Ser Asp Phe 245250 255 Ser Ser Ser Asp Ile Ile Asp Ser Ser Gln Lys Leu Gln Gln Asn Gly260 265 270 Phe Pro Glu Lys Val Glu Gln Met Asp Lys Tyr Ser Asn Lys LeuLys 275 280 285 Asp Glu Ala Ser Asp Lys Lys Tyr Glu Lys Pro Gly Lys LysAsp Tyr 290 295 300 Val Glu Glu Glu Gly Tyr Trp Ala Pro Ile Thr Asp SerGlu Asp Asp 305 310 315 320 Glu Ala 4 1108 DNA Caenorhabditis elegans 4gcaaaaaact agatattttg tggcattttt acaattaaaa aacctttaaa aaatggatca 60ccatgctatg taccgaaccg ctgaattcaa caaaactact gtccgattat tggcggaatt 120catcgaaaag actgggcaga atgcgacgat agtgaatatg gacagctttt tggagttctt 180tgcgtatttg aatcccacgg ctccaattcc aacggttcca gaaattgaaa aacaattatt 240gctaaaatca ccgattcgtt gcattgtgtg tggaatggaa actgaatcag attccgcagt 300gacattaagc atcgataatg cttcaattat tctcacagcg acagtaattg gttactgtag 360agatccaagt gatgcagtta atcaaattcg aaaggagagt cttcgagcat gcacgaaaca 420tttcaacagt attttccatg tcatcttcga aggactgcaa atcgagaaca cctactgtgc 480tcatcatgca aaatacagtc ttgccaatcg ttggtgcaaa gtctacacga tgattcgatc 540ttccctgggc gagcagttca caaagttcga tgtgcgcaat tttaaatcaa tattgcaatc 600atttttggat acttttggtg aaattgatga cgacaaaaag gataaagaat cttctcattt 660tgatgaatgt tttgaagaaa tggattcaga aaacgtagaa attaaaatgg agagcccaca 720agaagaagct gcagagaaat cgaagttttc tgaaaaccta gtggaggtaa aactggaacc 780aattgaaact catgaacttg acaaaactat atccgacttt tcttcaagtg atataattga 840ttcgtcccaa aaactgcagc aaaatggttt tcctgaaaaa gtggagcaaa tggacaaata 900tagcaacaaa ttgaaagatg aagcttcaga caaaaagtat gaaaagccag gaaaaaagga 960ctacgttgaa gaagagggat actgggcgcc gatcaccgac agcgaggatg atgaagcctg 1020aatttattta atcaaacgtt ttggaaattt tttttgtttt tgtcaataaa accatataac 1080aataaaaaaa aaaaaaaaaa aactcgag 1108 5 498 PRT Caenorhabditis elegans 5Met Ser Glu Phe Leu Lys Ile Val Arg Ala Asn Lys Lys Ser Asp Arg 1 5 1015 Lys Leu Asp Lys Thr Tyr Leu Trp Glu Ser Tyr Leu His Gln Phe Glu 20 2530 Lys Gly Lys Thr Ser Phe Ile Pro Val Glu Ala Phe Asn Arg Asn Leu 35 4045 Thr Val Asn Phe Asn Glu Cys Val Lys Glu Gly Val Ile Phe Glu Thr 50 5560 Val Val His Asp Tyr Asp Lys Asn Cys Asp Ser Ile Gln Val Arg Trp 65 7075 80 Phe Ala Arg Ile Glu Lys Val Cys Gly Tyr Arg Val Leu Ala Gln Phe 8590 95 Ile Gly Ala Asp Thr Lys Phe Trp Leu Asn Ile Leu Ser Asp Asp Met100 105 110 Phe Gly Leu Ala Asn Ala Ala Met Ser Asp Pro Asn Met Asp LysIle 115 120 125 Val Tyr Ala Pro Pro Leu Ala Ile Asn Glu Glu Tyr Gln AsnAsp Met 130 135 140 Val Asn Tyr Val Asn Asn Cys Ile Asp Gly Glu Ile ValGly Gln Thr 145 150 155 160 Ser Leu Ser Pro Lys Phe Asp Glu Gly Lys AlaLeu Leu Ser Lys His 165 170 175 Arg Phe Lys Val Gly Gln Arg Leu Glu LeuLeu Asn Tyr Ser Asn Ser 180 185 190 Thr Glu Ile Arg Val Ala Arg Ile GlnGlu Ile Cys Gly Arg Arg Met 195 200 205 Asn Val Ser Ile Thr Lys Lys AspPhe Pro Glu Ser Leu Pro Asp Ala 210 215 220 Asp Asp Asp Arg Gln Val PheSer Ser Gly Ser Gln Tyr Trp Ile Asp 225 230 235 240 Glu Gly Ser Phe PheIle Phe Pro Val Gly Phe Ala Ala Val Asn Gly 245 250 255 Tyr Gln Leu AsnAla Lys Lys Glu Tyr Ile Glu His Thr Asn Lys Ile 260 265 270 Ala Gln AlaIle Lys Asn Gly Glu Asn Pro Arg Tyr Asp Ser Asp Asp 275 280 285 Val ThrPhe Asp Gln Leu Ala Lys Asp Pro Ile Asp Pro Met Ile Trp 290 295 300 ArgLys Val Lys Val Gly Gln Lys Phe Glu Leu Ile Asp Pro Leu Ala 305 310 315320 Gln Gln Phe Asn Asn Leu His Val Ala Ser Ile Leu Lys Phe Cys Lys 325330 335 Thr Glu Gly Tyr Leu Ile Val Gly Met Asp Gly Pro Asp Ala Leu Glu340 345 350 Asp Ser Phe Pro Ile His Ile Asn Asn Thr Phe Met Phe Pro ValGly 355 360 365 Tyr Ala Glu Lys Tyr Asn Leu Glu Leu Val Pro Pro Asp GluPhe Lys 370 375 380 Gly Thr Phe Arg Trp Asp Glu Tyr Leu Glu Lys Glu SerAla Glu Thr 385 390 395 400 Leu Pro Leu Asp Leu Phe Lys Pro Met Pro SerGln Glu Arg Leu Asp 405 410 415 Lys Phe Lys Val Ile Leu Ile Ser Lys ArgVal Gly Leu Arg Leu Glu 420 425 430 Ala Ala Asp Met Cys Glu Asn Gln PheIle Cys Pro Ala Thr Val Lys 435 440 445 Ser Val His Gly Arg Leu Ile AsnVal Asn Phe Asp Gly Trp Asp Glu 450 455 460 Glu Phe Asp Glu Leu Tyr AspVal Asp Ser His Asp Ile Leu Pro Ile 465 470 475 480 Gly Trp Cys Glu AlaHis Ser Tyr Val Leu Gln Pro Pro Lys Lys Tyr 485 490 495 Asn Tyr 6 1497DNA Caenorhabditis elegans 6 atgtctgaat ttctgaaaat tgtcagagct aacaaaaaatcggacagaaa actcgataag 60 acctacttgt gggaatccta tttacatcag ttcgagaaaggaaaaacttc tttcattcca 120 gttgaagcat tcaatcgtaa ccttacagtt aattttaacgaatgcgtgaa ggaaggagtt 180 atcttcgaaa cagtggtcca tgattatgac aagaactgcgattcgattca agtcagatgg 240 tttgcacgaa ttgaaaaagt ttgcggatac agagttctggctcagtttat cggagctgac 300 acgaaatttt ggctcaatat tttatcggac gatatgtttggtttggcaaa cgccgcaatg 360 agtgatccca atatggataa aattgtatat gctccgccgcttgcaatcaa cgaagaatac 420 caaaatgata tggtaaatta tgtaaataat tgcattgatggcgaaatcgt cggccaaact 480 tcgctgtctc caaaattcga tgaagggaag gctctcctaagcaagcatcg tttcaaagtt 540 ggacaacgtc ttgaactatt aaattattcc aattctactgaaatacgcgt agcgcgaatt 600 caagaaatat gtggacgacg aatgaatgta tctatcacaaagaaagactt tcccgaatcg 660 cttccagatg cagatgacga cagacaagtc tttagctctggatctcaata ttggatagac 720 gagggaagct tcttcatatt tcctgttgga tttgcagcagtcaatggata tcaactaaat 780 gcgaaaaagg aatatattga gcacacaaat aaaattgctcaagcaataaa aaatggagaa 840 aatccaagat atgactcaga cgacgtcaca tttgatcaattagcaaaaga tccaattgat 900 cccatgattt ggagaaaagt taaggttgga caaaagtttgagctcatcga ccccttggct 960 cagcaattca ataacctcca cgtcgcttcg attctcaaattttgcaaaac tgaaggatat 1020 cttattgtgg gaatggatgg tccagatgca cttgaagacagttttcctat tcatatcaat 1080 aatacattta tgttcccagt tggttatgcg gaaaagtataatttggaact tgttccgcca 1140 gatgagttca aaggaacatt cagatgggat gaatacttggagaaagaatc tgcagaaacc 1200 ctaccgcttg acttgttcaa gccaatgcct tcccaagagagattagacaa atttaaggta 1260 attctgattt ccaaacgggt aggactacgc cttgaagctgctgacatgtg tgaaaatcag 1320 tttatttgtc cagctacagt gaaatcagtt catggaagactgataaatgt caatttcgac 1380 ggctgggatg aagaatttga tgaactgtat gatgtggactcccatgatat tctaccgata 1440 ggatggtgtg aagcgcacag ttatgttcta caacctccgaaaaagtacaa ctattga 1497 7 100 PRT Artificial Sequence derived fromCaenorhabditis elegans, Drosophila melanogaster, Mus musculus and Homosapiens 7 Phe Asp Trp Glu Asp Tyr Leu Glu Glu Thr Gly Ala Arg Ala AlaPro 1 5 10 15 Val Glu Leu Phe Asp Lys Gln Pro Val Asp Ser Pro Pro AsnGly Phe 20 25 30 Lys Val Gly Met Lys Leu Glu Ala Val Asp Pro Arg Asn ProSer Leu 35 40 45 Ile Cys Val Ala Thr Val Val Glu Val Lys Gly Tyr Arg LeuLeu Leu 50 55 60 His Phe Asp Gly Trp Asp Asp Arg Tyr Asp Phe Trp Cys AspAla Asp 65 70 75 80 Ser Pro Asp Ile Phe Pro Val Gly Trp Cys Glu Lys AsnGly His Pro 85 90 95 Leu Gln Pro Pro 100 8 100 PRT Drosophilamelanogaster 8 Trp Ser Trp Glu Ser Tyr Leu Glu Glu Gln Lys Ala Ile ThrAla Pro 1 5 10 15 Val Ser Leu Phe Asp Ser Gln Ala Val Thr His Asn LysAsn Gly Phe 20 25 30 Lys Leu Gly Met Lys Leu Glu Gly Ile Asp Pro Gln HisPro Ser Met 35 40 45 Tyr Phe Ile Leu Thr Val Ala Glu Val Cys Gly Tyr ArgLeu Arg Leu 50 55 60 His Phe Asp Gly Tyr Ser Glu Cys His Asp Phe Trp ValAsn Ala Asn 65 70 75 80 Ser Pro Asp Ile His Pro Ala Gly Trp Phe Glu LysThr Gly His Lys 85 90 95 Leu Gln Leu Pro 100 9 100 PRT Drosophilamelanogaster 9 Phe Ser Trp Ser Gln Tyr Met Cys Ser Thr Arg Ala Gln AlaAla Pro 1 5 10 15 Lys His Met Phe Val Ser Gln Ser His Ser Pro Pro ProLeu Gly Phe 20 25 30 Gln Val Gly Met Lys Leu Glu Ala Val Asp Arg Met AsnPro Ser Leu 35 40 45 Val Cys Val Ala Ser Val Thr Asp Val Val Asp Ser ArgPhe Leu Val 50 55 60 His Phe Asp Asn Trp Asp Asp Thr Tyr Asp Tyr Trp CysAsp Pro Ser 65 70 75 80 Ser Pro Tyr Ile His Pro Val Gly Trp Cys Gln LysGln Gly Lys Pro 85 90 95 Leu Thr Pro Pro 100 10 96 PRT Drosophilamelanogaster 10 Phe Cys Trp Glu Lys Tyr Leu Glu Glu Thr Gly Ala Ser AlaVal Pro 1 5 10 15 Thr Trp Ala Phe Lys Val Arg Pro Pro His Ser Phe LeuVal Asn Met 20 25 30 Lys Leu Glu Ala Val Asp Arg Arg Asn Pro Ala Leu IleArg Val Ala 35 40 45 Ser Val Glu Asp Val Glu Asp His Arg Ile Lys Ile HisPhe Asp Gly 50 55 60 Trp Ser His Gly Tyr Asp Phe Trp Ile Asp Ala Asp HisPro Asp Ile 65 70 75 80 His Pro Ala Gly Trp Cys Ser Lys Thr Gly His ProLeu Gln Pro Pro 85 90 95 11 99 PRT Drosophila melanogaster 11 Phe ArgTrp Ser Glu Tyr Leu Ser Lys Gly Lys Asp Val Ala Ala Pro 1 5 10 15 IleHis Leu Phe Leu Asn Pro Phe Pro Ile Ser Pro Asn Cys Phe Glu 20 25 30 IleGly Met Lys Leu Glu Ala Ile Asp Pro Glu Asn Cys Ser Leu Phe 35 40 45 CysVal Cys Ser Ile Val Glu Val Arg Gly Tyr Arg Leu Lys Leu Ser 50 55 60 PheAsp Gly Tyr Ser Ser Met Tyr Asp Phe Trp Val Asn Ala Asp Ser 65 70 75 80Gln Asp Ile Phe Pro Pro Gly Trp Cys Asp Glu Thr Ala Arg Val Leu 85 90 95Gln Ala Pro 12 100 PRT Drosophila melanogaster 12 Phe Ser Trp Ser ArgTyr Leu Val Lys Thr Gly Gly Lys Ala Ala Pro 1 5 10 15 Arg Ala Leu PheAsn Met Gln Gln Gln Met Asp Val Arg Asn Gly Phe 20 25 30 Ala Val Gly MetHis Leu Glu Ala Glu Asp Leu Asn Asp Thr Gly Lys 35 40 45 Ile Cys Val AlaThr Val Thr Asp Ile Leu Asp Glu Arg Ile Arg Val 50 55 60 His Phe Asp GlyTrp Asp Asp Cys Tyr Asp Leu Trp Val His Ile Thr 65 70 75 80 Ser Pro TyrIle His Pro Cys Gly Trp His Glu Gly Arg Gln Gln Leu 85 90 95 Ile Val ProPro 100 13 96 PRT Drosophila melanogaster 13 Phe Ile Trp Asp Asp Tyr IleSer Glu Val Gly Gly Met Ala Ala Ser 1 5 10 15 Lys Glu Leu Phe Thr ProArg Gln Pro Met Glu Tyr Gln Glu Arg Met 20 25 30 Lys Leu Glu Val Val AspGln Arg Asn Pro Cys Leu Ile Arg Pro Ala 35 40 45 Thr Val Val Thr Arg LysGly Tyr Arg Val Gln Leu His Leu Asp Cys 50 55 60 Trp Pro Thr Glu Tyr TyrPhe Trp Leu Glu Asp Asp Ser Pro Asp Leu 65 70 75 80 His Pro Ile Gly TrpCys Glu Ala Thr Ser His Glu Leu Glu Thr Pro 85 90 95 14 99 PRT Musmusculus 14 Phe Thr Trp Asp Lys Tyr Leu Lys Glu Thr Cys Ser Val Pro AlaPro 1 5 10 15 Val His Cys Phe Lys Gln Ser Tyr Thr Pro Pro Ser Asn GluPhe Lys 20 25 30 Ile Ser Met Lys Leu Glu Ala Gln Asp Pro Arg Asn Thr ThrSer Thr 35 40 45 Cys Ile Ala Thr Val Val Gly Leu Thr Gly Ala Arg Leu ArgLeu Arg 50 55 60 Leu Asp Gly Ser Asp Asn Lys Asn Asp Phe Trp Arg Leu ValAsp Ser 65 70 75 80 Ser Glu Ile Gln Pro Ile Gly Asn Cys Glu Lys Asn GlyGly Met Leu 85 90 95 Gln Pro Pro 15 100 PRT Homo sapiens 15 Ser Ser TrpPro Met Phe Leu Thr Leu Asn Gly Ser Glu Met Ala Ser 1 5 10 15 Ala ThrLeu Phe Lys Lys Glu Pro Pro Lys Pro Pro Leu Asn Asn Phe 20 25 30 Lys ValGly Met Lys Leu Glu Ala Ile Asp Lys Lys Asn Pro Tyr Leu 35 40 45 Ile CysPro Ala Thr Ile Gly Asp Val Lys Gly Asp Glu Val His Ile 50 55 60 Thr PheAsp Gly Trp Ser Gly Ala Phe Asp Tyr Trp Cys Lys Tyr Asp 65 70 75 80 SerArg Asp Ile Phe Pro Ala Gly Trp Cys Arg Leu Thr Gly Asp Val 85 90 95 LeuGln Pro Pro 100 16 1276 DNA Caenorhabditis elegans 16 agaatctgccaaaatgtcaa agataaagac acattccact ggctcaaaac ggacggtacc 60 attctacaagccgccaccgc ccgtgccact tccaccactc ccgccacccg atccaacccg 120 gtacttctcgacggaaaagt acatcgcact gagcaaagat gagaaattca aatttgatga 180 ttacgatgtgaatgatgaga cgctgaaaaa agtggtgctc aacgagattg gcaagtgccc 240 ggatatttggagctcgcgga gccaggcagc cattatggag cactatccga ttgttgcaac 300 tgaaacgtacaggaggacag ggttgctgtt gtctatcaaa tcgctgaaac aaatctacaa 360 atgcggaaaggacaatctcc gaaaccggct tcgcgtggca attgtaagca agcggcttac 420 accggcccaagtagaggcct atatgtggcg ctgggagttt tacggcttta ttcgctacta 480 tcgagactatacacaacgct gggaggccga cttgttgaaa gatttggacg tggtgctcgg 540 gctcgaggctcggcgagcat cgaaaaatat ggaaaaggtg gattctgggg agctcatgga 600 gcccatggagcccatggatt ctacaatgga tgagatgtgc gtcgaggagg agccctacga 660 ggagacagggtccaattgga gcgatccggc gccggaacca tcccaatcca aatcccagtc 720 cccagaagccaagtaccctc aagcctacct actacctgag gcggacgaag tctacaatcc 780 tgacgatttctatcaagagg aacatgaatc cgcatcaaac gccatgtatc ggatcgcttt 840 ctcacagcagtacggtggcg gcgggtcccc agccgtgcag aagcccgtca cttttagtgc 900 tcagccggcgccggcgccag ttagagaggc cccaagccca gttgtggaga atgttagttc 960 atcgagtttcaccccgaagc ccccggccat gatcaacaat tttggtgagg agatgaacca 1020 aataacataccaagcgatcc gtattgcccg agagcagccg gaacgtctga aattgctccg 1080 taaggcacttttcgacgttg tcctggcgtt tgatcagaag gaatacgccg atgttgggga 1140 tttgtacagggatttggcgc aaaagaattc gtgataattt ttttttgagt tttttaattt 1200 ttaatttatttcaatttttg ttacatgttc caatataata aacaggtgct tgtttaaaaa 1260 aaaaaaaaaaaaaaaa 1276 17 386 PRT Caenorhabditis elegans 17 Met Ser Lys Ile Lys ThrHis Ser Thr Gly Ser Lys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Pro ProPro Pro Val Pro Leu Pro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg TyrPhe Ser Thr Glu Lys Tyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe LysPhe Asp Asp Tyr Asp Val Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val LeuAsn Glu Ile Gly Lys Cys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser GlnAla Ala Ile Met Glu His Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr ArgArg Thr Gly Leu Leu Leu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile TyrLys Cys Gly Lys Asp Asn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala IleVal Ser Lys Arg Leu Thr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 TrpArg Trp Glu Phe Tyr Gly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155160 Gln Arg Trp Glu Ala Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165170 175 Leu Glu Ala Arg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly180 185 190 Glu Leu Met Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp GluMet 195 200 205 Cys Val Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn TrpSer Asp 210 215 220 Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser ProGlu Ala Lys 225 230 235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala AspGlu Val Tyr Asn Pro 245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu SerAla Ser Asn Ala Met Tyr 260 265 270 Arg Ile Ala Phe Ser Gln Gln Tyr GlyGly Gly Gly Ser Pro Ala Val 275 280 285 Gln Lys Pro Val Thr Phe Ser AlaGln Pro Ala Pro Ala Pro Val Arg 290 295 300 Glu Ala Pro Ser Pro Val ValGlu Asn Val Ser Ser Ser Ser Phe Thr 305 310 315 320 Pro Lys Pro Pro AlaMet Ile Asn Asn Phe Gly Glu Glu Met Asn Gln 325 330 335 Ile Thr Tyr GlnAla Ile Arg Ile Ala Arg Glu Gln Pro Glu Arg Leu 340 345 350 Lys Leu LeuArg Lys Ala Leu Phe Asp Val Val Leu Ala Phe Asp Gln 355 360 365 Lys GluTyr Ala Asp Val Gly Asp Leu Tyr Arg Asp Leu Ala Gln Lys 370 375 380 AsnSer 385 18 1276 DNA Caenorhabditis elegans 18 agaatctgcc aaaatgtcaaagataaagac acattccact ggctcaaaac ggacggtacc 60 attctacaag ctgccaccgcccgtgccact tccaccactc ccgccacccg atccaacccg 120 gtacttctcg acggaaaagtacatcgcact gagcaaagat gagaaattca aatttgatga 180 ttacgatgtg aatgatgagacgctgaaaaa agtgatgctc aacgagattg gcaagtgccc 240 ggatatttgg agctcgcggagccaggcagc cattatggag cactatccga ttgttgcaac 300 tgaaacgtac aggaggacagggttgctgtt gtctatcaaa tcgctgaaac aaatctacaa 360 atgcggaaag gacaatctccgaaaccggct tcgcgtggca attgtaagca agcggcttac 420 accggcccaa gtagaggcctatatgtggcg ctgggagttt tacggcttta ttcgctacta 480 tcgagactat acacaacgctgggaggccga cttgttgaaa gatttggacg tggtgctcgg 540 gctcgaggct cggcgagcatcgaaaaatat ggaaaaggtg gattctgggg agctcatgga 600 gcccatggag cccatggattctacaatgga tgagatgtgc gtcgaggagg agccctacga 660 ggagacaggg tccaattggagcgatccggc gccggaacca tcccaatcca aatcccagtc 720 cccagaagcc aagtaccctcaagcctacct actacctgag gcggacgaag tctacaatcc 780 tgacgatttc tatcaagaggaacatgaatc cgcatcaaac gccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcggcgggtcccc agccgtgcag aagcccgtca cttttagtgc 900 tcagccggcg ccggcgccagttagagaggc cccaagccca gttgtggaga atgttagttc 960 atcgagtttc accccgaagcccccggccat gatcaacaat tttggtgagg agatgaacca 1020 aataacatac caagcgatccgtattgcccg agagcagccg gaacgtctga aattgctccg 1080 taaggcactt ttcgacgttgtcctggcgtt tgatcagaag gaatacgccg atgttgggga 1140 tttgtacagg gatttggcgcaaaagaattc gtgataattt ttttttgagt tttttaattt 1200 ttaatttatt tcaatttttgttacatgttc caatataata aacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 127619 386 PRT Caenorhabditis elegans 19 Met Ser Lys Ile Lys Thr His Ser ThrGly Ser Lys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro ValPro Leu Pro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser ThrGlu Lys Tyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp AspTyr Asp Val Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Met Leu Asn Glu IleGly Lys Cys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala IleMet Glu His Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr GlyLeu Leu Leu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys GlyLys Asp Asn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser LysArg Leu Thr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg Trp GluPhe Tyr Gly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln ArgTrp Glu Ala Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175 LeuGlu Ala Arg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly 180 185 190Glu Leu Met Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp Glu Met 195 200205 Cys Val Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn Trp Ser Asp 210215 220 Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser Pro Glu Ala Lys225 230 235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala Asp Glu Val TyrAsn Pro 245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu Ser Ala Ser AsnAla Met Tyr 260 265 270 Arg Ile Ala Phe Ser Gln Gln Tyr Gly Gly Gly GlySer Pro Ala Val 275 280 285 Gln Lys Pro Val Thr Phe Ser Ala Gln Pro AlaPro Ala Pro Val Arg 290 295 300 Glu Ala Pro Ser Pro Val Val Glu Asn ValSer Ser Ser Ser Phe Thr 305 310 315 320 Pro Lys Pro Pro Ala Met Ile AsnAsn Phe Gly Glu Glu Met Asn Gln 325 330 335 Ile Thr Tyr Gln Ala Ile ArgIle Ala Arg Glu Gln Pro Glu Arg Leu 340 345 350 Lys Leu Leu Arg Lys AlaLeu Phe Asp Val Val Leu Ala Phe Asp Gln 355 360 365 Lys Glu Tyr Ala AspVal Gly Asp Leu Tyr Arg Asp Leu Ala Gln Lys 370 375 380 Asn Ser 385 201276 DNA Caenorhabditis elegans 20 agaatctgcc aaaatgtcaa agataaagacacattccact ggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccacttccaccactc ccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcactgagcaaagat gagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaaagtggtgctc aacgagattg gcaagtgccc 240 ggatatttag agctcgcgga gccaggcagccattatggag cactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgttgtctatcaaa tcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggcttcgcgtggca attgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcgctgggagttt tacggcttta ttcgctacta 480 tcgagactat acacaacgct gggaggccgacttgttgaaa gatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatatggaaaaggtg gattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatggatgagatgtgc gtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggcgccggaacca tcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacctactacctgag gcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatccgcatcaaac gccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtccccagccgtgcag aagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggccccaagccca gttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccatgatcaacaat tttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccgagagcagccg gaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtttgatcagaag gaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattcgtgataattt ttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttccaatataata aacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 21 78 PRTCaenorhabditis elegans 21 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile 65 70 75 22 1276 DNA Caenorhabditis elegans 22agaatctgcc aaaatgtcaa agataaagac acattccact ggctcaaaac ggacggtacc 60attctacaag ctgccaccgc ccgtgccact tccaccactc ccgccacccg atccaacccg 120gtacttctcg acggaaaagt acatcgcact gagcaaagat gagaaattca aatttgatga 180ttacgatgtg aatgatgaga cgctgaaaaa agtggtgctc aacgagattg gcaagtgccc 240ggatatttgg agctcgcgga gccaggcagc cattatggag cactatccga ttgttgcaac 300tgaaacgtac aggaggacag ggttgctgtt gtctatcaaa tcgctgaaat aaatctacaa 360atgcggaaag gacaatctcc gaaaccggct tcgcgtggca attgtaagca agcggcttac 420accggcccaa gtagaggcct atatgtggcg ctgggagttt tacggcttta ttcgctacta 480tcgagactat acacaacgct gggaggccga cttgttgaaa gatttggacg tggtgctcgg 540gctcgaggct cggcgagcat cgaaaaatat ggaaaaggtg gattctgggg agctcatgga 600gcccatggag cccatggatt ctacaatgga tgagatgtgc gtcgaggagg agccctacga 660ggagacaggg tccaattgga gcgatccggc gccggaacca tcccaatcca aatcccagtc 720cccagaagcc aagtaccctc aagcctacct actacctgag gcggacgaag tctacaatcc 780tgacgatttc tatcaagagg aacatgaatc cgcatcaaac gccatgtatc ggatcgcttt 840ctcacagcag tacggtggcg gcgggtcccc agccgtgcag aagcccgtca cttttagtgc 900tcagccggcg ccggcgccag ttagagaggc cccaagccca gttgtggaga atgttagttc 960atcgagtttc accccgaagc ccccggccat gatcaacaat tttggtgagg agatgaacca 1020aataacatac caagcgatcc gtattgcccg agagcagccg gaacgtctga aattgctccg 1080taaggcactt ttcgacgttg tcctggcgtt tgatcagaag gaatacgccg atgttgggga 1140tttgtacagg gatttggcgc aaaagaattc gtgataattt ttttttgagt tttttaattt 1200ttaatttatt tcaatttttg ttacatgttc caatataata aacaggtgct tgtttaaaaa 1260aaaaaaaaaa aaaaaa 1276 23 112 PRT Caenorhabditis elegans 23 Met Ser LysIle Lys Thr His Ser Thr Gly Ser Lys Arg Thr Val Pro 1 5 10 15 Phe TyrLys Leu Pro Pro Pro Val Pro Leu Pro Pro Leu Pro Pro Pro 20 25 30 Asp ProThr Arg Tyr Phe Ser Thr Glu Lys Tyr Ile Ala Leu Ser Lys 35 40 45 Asp GluLys Phe Lys Phe Asp Asp Tyr Asp Val Asn Asp Glu Thr Leu 50 55 60 Lys LysVal Val Leu Asn Glu Ile Gly Lys Cys Pro Asp Ile Trp Ser 65 70 75 80 SerArg Ser Gln Ala Ala Ile Met Glu His Tyr Pro Ile Val Ala Thr 85 90 95 GluThr Tyr Arg Arg Thr Gly Leu Leu Leu Ser Ile Lys Ser Leu Lys 100 105 11024 1276 DNA Caenorhabditis elegans 24 agaatctgcc aaaatgtcaa agataaagacacattccact ggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccacttccaccactc ccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcactgagcaaagat gagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaaagtggtgctc aacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagccattatggag cactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgttgtctatcaaa tcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggcttcacgtggca attgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcgctgggagttt tacggcttta ttcgctacta 480 tcgagactat acacaacgct gggaggccgacttgttgaaa gatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatatggaaaaggtg gattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatggatgagatgtgc gtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggcgccggaacca tcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacctactacctgag gcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatccgcatcaaac gccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtccccagccgtgcag aagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggccccaagccca gttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccatgatcaacaat tttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccgagagcagccg gaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtttgatcagaag gaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattcgtgataattt ttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttccaatataata aacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 25 386 PRTCaenorhabditis elegans 25 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu His Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg Trp Glu Phe TyrGly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln Arg Trp GluAla Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175 Leu Glu AlaArg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly 180 185 190 Glu LeuMet Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp Glu Met 195 200 205 CysVal Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn Trp Ser Asp 210 215 220Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser Pro Glu Ala Lys 225 230235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala Asp Glu Val Tyr Asn Pro245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu Ser Ala Ser Asn Ala MetTyr 260 265 270 Arg Ile Ala Phe Ser Gln Gln Tyr Gly Gly Gly Gly Ser ProAla Val 275 280 285 Gln Lys Pro Val Thr Phe Ser Ala Gln Pro Ala Pro AlaPro Val Arg 290 295 300 Glu Ala Pro Ser Pro Val Val Glu Asn Val Ser SerSer Ser Phe Thr 305 310 315 320 Pro Lys Pro Pro Ala Met Ile Asn Asn PheGly Glu Glu Met Asn Gln 325 330 335 Ile Thr Tyr Gln Ala Ile Arg Ile AlaArg Glu Gln Pro Glu Arg Leu 340 345 350 Lys Leu Leu Arg Lys Ala Leu PheAsp Val Val Leu Ala Phe Asp Gln 355 360 365 Lys Glu Tyr Ala Asp Val GlyAsp Leu Tyr Arg Asp Leu Ala Gln Lys 370 375 380 Asn Ser 385 26 1276 DNACaenorhabditis elegans 26 agaatctgcc aaaatgtcaa agataaagac acattccactggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccact tccaccactcccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcact gagcaaagatgagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaa agtggtgctcaacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagc cattatggagcactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgtt gtctatcaaatcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggct tcgcgtggcaattgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggca ctgggagttttacggcttta ttcgctacta 480 tcgagactat acacaacgct gggaggccga cttgttgaaagatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatat ggaaaaggtggattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatgga tgagatgtgcgtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggc gccggaaccatcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacct actacctgaggcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatc cgcatcaaacgccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtcccc agccgtgcagaagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggc cccaagcccagttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccat gatcaacaattttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccg agagcagccggaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtt tgatcagaaggaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattc gtgataatttttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttc caatataataaacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 27 386 PRTCaenorhabditis elegans 27 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp His Trp Glu Phe TyrGly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln Arg Trp GluAla Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175 Leu Glu AlaArg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly 180 185 190 Glu LeuMet Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp Glu Met 195 200 205 CysVal Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn Trp Ser Asp 210 215 220Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser Pro Glu Ala Lys 225 230235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala Asp Glu Val Tyr Asn Pro245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu Ser Ala Ser Asn Ala MetTyr 260 265 270 Arg Ile Ala Phe Ser Gln Gln Tyr Gly Gly Gly Gly Ser ProAla Val 275 280 285 Gln Lys Pro Val Thr Phe Ser Ala Gln Pro Ala Pro AlaPro Val Arg 290 295 300 Glu Ala Pro Ser Pro Val Val Glu Asn Val Ser SerSer Ser Phe Thr 305 310 315 320 Pro Lys Pro Pro Ala Met Ile Asn Asn PheGly Glu Glu Met Asn Gln 325 330 335 Ile Thr Tyr Gln Ala Ile Arg Ile AlaArg Glu Gln Pro Glu Arg Leu 340 345 350 Lys Leu Leu Arg Lys Ala Leu PheAsp Val Val Leu Ala Phe Asp Gln 355 360 365 Lys Glu Tyr Ala Asp Val GlyAsp Leu Tyr Arg Asp Leu Ala Gln Lys 370 375 380 Asn Ser 385 28 1276 DNACaenorhabditis elegans 28 agaatctgcc aaaatgtcaa agataaagac acattccactggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccact tccaccactcccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcact gagcaaagatgagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaa agtggtgctcaacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagc cattatggagcactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgtt gtctatcaaatcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggct tcgcgtggcaattgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcg ctgagagttttacggcttta ttcgctacta 480 tcgagactat acacaacgct gggaggccga cttgttgaaagatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatat ggaaaaggtggattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatgga tgagatgtgcgtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggc gccggaaccatcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacct actacctgaggcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatc cgcatcaaacgccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtcccc agccgtgcagaagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggc cccaagcccagttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccat gatcaacaattttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccg agagcagccggaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtt tgatcagaaggaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattc gtgataatttttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttc caatataataaacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 29 146 PRTCaenorhabditis elegans 29 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg 145 30 1276 DNACaenorhabditis elegans 30 agaatctgcc aaaatgtcaa agataaagac acattccactggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccact tccaccactcccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcact gagcaaagatgagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaa agtggtgctcaacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagc cattatggagcactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgtt gtctatcaaatcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggct tcgcgtggcaattgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcg ctggaagttttacggcttta ttcgctacta 480 tcgagactat acacaacgct gggaggccga cttgttgaaagatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatat ggaaaaggtggattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatgga tgagatgtgcgtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggc gccggaaccatcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacct actacctgaggcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatc cgcatcaaacgccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtcccc agccgtgcagaagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggc cccaagcccagttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccat gatcaacaattttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccg agagcagccggaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtt tgatcagaaggaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattc gtgataatttttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttc caatataataaacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 31 386 PRTCaenorhabditis elegans 31 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg Trp Lys Phe TyrGly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln Arg Trp GluAla Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175 Leu Glu AlaArg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly 180 185 190 Glu LeuMet Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp Glu Met 195 200 205 CysVal Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn Trp Ser Asp 210 215 220Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser Pro Glu Ala Lys 225 230235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala Asp Glu Val Tyr Asn Pro245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu Ser Ala Ser Asn Ala MetTyr 260 265 270 Arg Ile Ala Phe Ser Gln Gln Tyr Gly Gly Gly Gly Ser ProAla Val 275 280 285 Gln Lys Pro Val Thr Phe Ser Ala Gln Pro Ala Pro AlaPro Val Arg 290 295 300 Glu Ala Pro Ser Pro Val Val Glu Asn Val Ser SerSer Ser Phe Thr 305 310 315 320 Pro Lys Pro Pro Ala Met Ile Asn Asn PheGly Glu Glu Met Asn Gln 325 330 335 Ile Thr Tyr Gln Ala Ile Arg Ile AlaArg Glu Gln Pro Glu Arg Leu 340 345 350 Lys Leu Leu Arg Lys Ala Leu PheAsp Val Val Leu Ala Phe Asp Gln 355 360 365 Lys Glu Tyr Ala Asp Val GlyAsp Leu Tyr Arg Asp Leu Ala Gln Lys 370 375 380 Asn Ser 385 32 1276 DNACaenorhabditis elegans 32 agaatctgcc aaaatgtcaa agataaagac acattccactggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccact tccaccactcccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcact gagcaaagatgagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaa agtggtgctcaacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagc cattatggagcactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgtt gtctatcaaatcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggct tcgcgtggcaattgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcg ctgggagttttacggcttta tttgctacta 480 tcgagactat acacaacgct gggaggccga cttgttgaaagatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatat ggaaaaggtggattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatgga tgagatgtgcgtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggc gccggaaccatcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacct actacctgaggcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatc cgcatcaaacgccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtcccc agccgtgcagaagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggc cccaagcccagttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccat gatcaacaattttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccg agagcagccggaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtt tgatcagaaggaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattc gtgataatttttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttc caatataataaacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 33 386 PRTCaenorhabditis elegans 33 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg Trp Glu Phe TyrGly Phe Ile Cys Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln Arg Trp GluAla Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175 Leu Glu AlaArg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly 180 185 190 Glu LeuMet Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp Glu Met 195 200 205 CysVal Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn Trp Ser Asp 210 215 220Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser Pro Glu Ala Lys 225 230235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala Asp Glu Val Tyr Asn Pro245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu Ser Ala Ser Asn Ala MetTyr 260 265 270 Arg Ile Ala Phe Ser Gln Gln Tyr Gly Gly Gly Gly Ser ProAla Val 275 280 285 Gln Lys Pro Val Thr Phe Ser Ala Gln Pro Ala Pro AlaPro Val Arg 290 295 300 Glu Ala Pro Ser Pro Val Val Glu Asn Val Ser SerSer Ser Phe Thr 305 310 315 320 Pro Lys Pro Pro Ala Met Ile Asn Asn PheGly Glu Glu Met Asn Gln 325 330 335 Ile Thr Tyr Gln Ala Ile Arg Ile AlaArg Glu Gln Pro Glu Arg Leu 340 345 350 Lys Leu Leu Arg Lys Ala Leu PheAsp Val Val Leu Ala Phe Asp Gln 355 360 365 Lys Glu Tyr Ala Asp Val GlyAsp Leu Tyr Arg Asp Leu Ala Gln Lys 370 375 380 Asn Ser 385 34 1276 DNACaenorhabditis elegans 34 agaatctgcc aaaatgtcaa agataaagac acattccactggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccact tccaccactcccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcact gagcaaagatgagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaa agtggtgctcaacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagc cattatggagcactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgtt gtctatcaaatcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggct tcgcgtggcaattgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcg ctgggagttttacggcttta ttcgctacta 480 tcgagactat acacaacgct aggaggccga cttgttgaaagatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatat ggaaaaggtggattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatgga tgagatgtgcgtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggc gccggaaccatcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacct actacctgaggcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatc cgcatcaaacgccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtcccc agccgtgcagaagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggc cccaagcccagttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccat gatcaacaattttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccg agagcagccggaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtt tgatcagaaggaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattc gtgataatttttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttc caatataataaacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 35 162 PRTCaenorhabditis elegans 35 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg Trp Glu Phe TyrGly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln Arg 36 1276DNA Caenorhabditis elegans 36 agaatctgcc aaaatgtcaa agataaagacacattccact ggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccacttccaccactc ccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcactgagcaaagat gagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaaagtggtgctc aacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagccattatggag cactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgttgtctatcaaa tcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggcttcgcgtggca attgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcgctgggagttt tacggcttta ttcgctacta 480 tcgagactat acacaacgct ggaaggccgacttgttgaaa gatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatatggaaaaggtg gattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatggatgagatgtgc gtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggcgccggaacca tcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacctactacctgag gcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatccgcatcaaac gccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtccccagccgtgcag aagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggccccaagccca gttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccatgatcaacaat tttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccgagagcagccg gaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtttgatcagaag gaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattcgtgataattt ttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttccaatataata aacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 37 386 PRTCaenorhabditis elegans 37 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg Trp Glu Phe TyrGly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln Arg Trp LysAla Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175 Leu Glu AlaArg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly 180 185 190 Glu LeuMet Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp Glu Met 195 200 205 CysVal Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn Trp Ser Asp 210 215 220Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser Pro Glu Ala Lys 225 230235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala Asp Glu Val Tyr Asn Pro245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu Ser Ala Ser Asn Ala MetTyr 260 265 270 Arg Ile Ala Phe Ser Gln Gln Tyr Gly Gly Gly Gly Ser ProAla Val 275 280 285 Gln Lys Pro Val Thr Phe Ser Ala Gln Pro Ala Pro AlaPro Val Arg 290 295 300 Glu Ala Pro Ser Pro Val Val Glu Asn Val Ser SerSer Ser Phe Thr 305 310 315 320 Pro Lys Pro Pro Ala Met Ile Asn Asn PheGly Glu Glu Met Asn Gln 325 330 335 Ile Thr Tyr Gln Ala Ile Arg Ile AlaArg Glu Gln Pro Glu Arg Leu 340 345 350 Lys Leu Leu Arg Lys Ala Leu PheAsp Val Val Leu Ala Phe Asp Gln 355 360 365 Lys Glu Tyr Ala Asp Val GlyAsp Leu Tyr Arg Asp Leu Ala Gln Lys 370 375 380 Asn Ser 385 38 1275 DNACaenorhabditis elegans 38 agaatctgcc aaaatgtcaa agataaagac acattccactggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccact tccaccactcccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcact gagcaaagatgagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaa agtggtgctcaacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagc cattatggagcactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgtt gtctatcaaatcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggct tcgcgtggcaattgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcg ctgggagttttacggcttta ttcgctacta 480 tcgagactat acacaacgct gggaggccga cttgttgaaagatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatat ggaaaagtggattctgggga gctcatggag 600 cccatggagc ccatggattc tacaatggat gagatgtgcgtcgaggagga gccctacgag 660 gagacagggt ccaattggag cgatccggcg ccggaaccatcccaatccaa atcccagtcc 720 ccagaagcca agtaccctca agcctaccta ctacctgaggcggacgaagt ctacaatcct 780 gacgatttct atcaagagga acatgaatcc gcatcaaacgccatgtatcg gatcgctttc 840 tcacagcagt acggtggcgg cgggtcccca gccgtgcagaagcccgtcac ttttagtgct 900 cagccggcgc cggcgccagt tagagaggcc ccaagcccagttgtggagaa tgttagttca 960 tcgagtttca ccccgaagcc cccggccatg atcaacaattttggtgagga gatgaaccaa 1020 ataacatacc aagcgatccg tattgcccga gagcagccggaacgtctgaa attgctccgt 1080 aaggcacttt tcgacgttgt cctggcgttt gatcagaaggaatacgccga tgttggggat 1140 ttgtacaggg atttggcgca aaagaattcg tgataatttttttttgagtt ttttaatttt 1200 taatttattt caatttttgt tacatgttcc aatataataaacaggtgctt gtttaaaaaa 1260 aaaaaaaaaa aaaaa 1275 39 325 PRTCaenorhabditis elegans 39 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg Trp Glu Phe TyrGly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln Arg Trp GluAla Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175 Leu Glu AlaArg Arg Ala Ser Lys Asn Met Glu Lys Trp Ile Leu Gly 180 185 190 Ser SerTrp Ser Pro Trp Ser Pro Trp Ile Leu Gln Trp Met Arg Cys 195 200 205 AlaSer Arg Arg Ser Pro Thr Arg Arg Gln Gly Pro Ile Gly Ala Ile 210 215 220Arg Arg Arg Asn His Pro Asn Pro Asn Pro Ser Pro Gln Lys Pro Ser 225 230235 240 Thr Leu Lys Pro Thr Tyr Tyr Leu Arg Arg Thr Lys Ser Thr Ile Leu245 250 255 Thr Ile Ser Ile Lys Arg Asn Met Asn Pro His Gln Thr Pro CysIle 260 265 270 Gly Ser Leu Ser His Ser Ser Thr Val Ala Ala Gly Pro GlnPro Cys 275 280 285 Arg Ser Pro Ser Leu Leu Val Leu Ser Arg Arg Arg ArgGln Leu Glu 290 295 300 Arg Pro Gln Ala Gln Leu Trp Arg Met Leu Val HisArg Val Ser Pro 305 310 315 320 Arg Ser Pro Arg Pro 325 40 1276 DNACaenorhabditis elegans 40 agaatctgcc aaaatgtcaa agataaagac acattccactggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccact tccaccactcccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcact gagcaaagatgagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaa agtggtgctcaacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagc cattatggagcactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgtt gtctatcaaatcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggct tcgcgtggcaattgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcg ctgggagttttacggcttta ttcgctacta 480 tcgagactat acacaacgct gggaggccga cttgttgaaagatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatat ggaaaaggtggattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatgga tgagatgtgcgtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggc gccggaaccatcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacct actacctgaggcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatc cgcatcaaacgccatgtatc ggatcgcttt 840 ctcacagtag tacggtggcg gcgggtcccc agccgtgcagaagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggc cccaagcccagttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccat gatcaacaattttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccg agagcagccggaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtt tgatcagaaggaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattc gtgataatttttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttc caatataataaacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 41 278 PRTCaenorhabditis elegans 41 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg Trp Glu Phe TyrGly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln Arg Trp GluAla Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175 Leu Glu AlaArg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly 180 185 190 Glu LeuMet Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp Glu Met 195 200 205 CysVal Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn Trp Ser Asp 210 215 220Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser Pro Glu Ala Lys 225 230235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala Asp Glu Val Tyr Asn Pro245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu Ser Ala Ser Asn Ala MetTyr 260 265 270 Arg Ile Ala Phe Ser Gln 275 42 1276 DNA Caenorhabditiselegans 42 agaatctgcc aaaatgtcaa agataaagac acattccact ggctcaaaacggacggtacc 60 attctacaag ctgccaccgc ccgtgccact tccaccactc ccgccacccgatccaacccg 120 gtacttctcg acggaaaagt acatcgcact gagcaaagat gagaaattcaaatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaa agtggtgctc aacgagattggcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagc cattatggag cactatccgattgttgcaac 300 tgaaacgtac aggaggacag ggttgctgtt gtctatcaaa tcgctgaaacaaatctacaa 360 atgcggaaag gacaatctcc gaaaccggct tcgcgtggca attgtaagcaagcggcttac 420 accggcccaa gtagaggcct atatgtggcg ctgggagttt tacggctttattcgctacta 480 tcgagactat acacaacgct gggaggccga cttgttgaaa gatttggacgtggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatat ggaaaaggtg gattctggggagctcatgga 600 gcccatggag cccatggatt ctacaatgga tgagatgtgc gtcgaggaggagccctacga 660 ggagacaggg tccaattgga gcgatccggc gccggaacca tcccaatccaaatcccagtc 720 cccagaagcc aagtaccctc aagcctacct actacctgag gcggacgaagtctacaatcc 780 tgacgatttc tatcaagagg aacatgaatc cgcatcaaac gccatgtatcggatcgcttt 840 ctcacagcag tacggtggcg gcgggtcccc agccgtgcag aagcccgtcacttttagtgc 900 tcagccggcg ccggcgccag tttgagaggc cccaagccca gttgtggagaatgttagttc 960 atcgagtttc accccgaagc ccccggccat gatcaacaat tttggtgaggagatgaacca 1020 aataacatac caagcgatcc gtattgcccg agagcagccg gaacgtctgaaattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtt tgatcagaag gaatacgccgatgttgggga 1140 tttgtacagg gatttggcgc aaaagaattc gtgataattt ttttttgagttttttaattt 1200 ttaatttatt tcaatttttg ttacatgttc caatataata aacaggtgcttgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 43 303 PRT Caenorhabditis elegans43 Met Ser Lys Ile Lys Thr His Ser Thr Gly Ser Lys Arg Thr Val Pro 1 510 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro Leu Pro Pro Leu Pro Pro Pro 2025 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu Lys Tyr Ile Ala Leu Ser Lys 3540 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr Asp Val Asn Asp Glu Thr Leu 5055 60 Lys Lys Val Val Leu Asn Glu Ile Gly Lys Cys Pro Asp Ile Trp Ser 6570 75 80 Ser Arg Ser Gln Ala Ala Ile Met Glu His Tyr Pro Ile Val Ala Thr85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu Leu Leu Ser Ile Lys Ser Leu Lys100 105 110 Gln Ile Tyr Lys Cys Gly Lys Asp Asn Leu Arg Asn Arg Leu ArgVal 115 120 125 Ala Ile Val Ser Lys Arg Leu Thr Pro Ala Gln Val Glu AlaTyr Met 130 135 140 Trp Arg Trp Glu Phe Tyr Gly Phe Ile Arg Tyr Tyr ArgAsp Tyr Thr 145 150 155 160 Gln Arg Trp Glu Ala Asp Leu Leu Lys Asp LeuAsp Val Val Leu Gly 165 170 175 Leu Glu Ala Arg Arg Ala Ser Lys Asn MetGlu Lys Val Asp Ser Gly 180 185 190 Glu Leu Met Glu Pro Met Glu Pro MetAsp Ser Thr Met Asp Glu Met 195 200 205 Cys Val Glu Glu Glu Pro Tyr GluGlu Thr Gly Ser Asn Trp Ser Asp 210 215 220 Pro Ala Pro Glu Pro Ser GlnSer Lys Ser Gln Ser Pro Glu Ala Lys 225 230 235 240 Tyr Pro Gln Ala TyrLeu Leu Pro Glu Ala Asp Glu Val Tyr Asn Pro 245 250 255 Asp Asp Phe TyrGln Glu Glu His Glu Ser Ala Ser Asn Ala Met Tyr 260 265 270 Arg Ile AlaPhe Ser Gln Gln Tyr Gly Gly Gly Gly Ser Pro Ala Val 275 280 285 Gln LysPro Val Thr Phe Ser Ala Gln Pro Ala Pro Ala Pro Val 290 295 300 44 1276DNA Caenorhabditis elegans 44 agaatctgcc aaaatgtcaa agataaagacacattccact ggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccacttccaccactc ccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcactgagcaaagat gagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaaagtggtgctc aacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagccattatggag cactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgttgtctatcaaa tcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggcttcgcgtggca attgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcgctgggagttt tacggcttta ttcgctacta 480 tcgagactat acacaacgct gggaggccgacttgttgaaa gatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatatggaaaaggtg gattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatggatgagatgtgc gtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggcgccggaacca tcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacctactacctgag gcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatccgcatcaaac gccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtccccagccgtgcag aagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggccccaagccca gttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccatgatcaacaat tttggtgagg agatgaacca 1020 aataacatac taagcgatcc gtattgcccgagagcagccg gaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtttgatcagaag gaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattcgtgataattt ttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttccaatataata aacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 45 339 PRTCaenorhabditis elegans 45 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg Trp Glu Phe TyrGly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln Arg Trp GluAla Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175 Leu Glu AlaArg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly 180 185 190 Glu LeuMet Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp Glu Met 195 200 205 CysVal Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn Trp Ser Asp 210 215 220Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser Pro Glu Ala Lys 225 230235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala Asp Glu Val Tyr Asn Pro245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu Ser Ala Ser Asn Ala MetTyr 260 265 270 Arg Ile Ala Phe Ser Gln Gln Tyr Gly Gly Gly Gly Ser ProAla Val 275 280 285 Gln Lys Pro Val Thr Phe Ser Ala Gln Pro Ala Pro AlaPro Val Arg 290 295 300 Glu Ala Pro Ser Pro Val Val Glu Asn Val Ser SerSer Ser Phe Thr 305 310 315 320 Pro Lys Pro Pro Ala Met Ile Asn Asn PheGly Glu Glu Met Asn Gln 325 330 335 Ile Thr Tyr 46 1276 DNACaenorhabditis elegans 46 agaatctgcc aaaatgtcaa agataaagac acattccactggctcaaaac ggacggtacc 60 attctacaag ctgccaccgc ccgtgccact tccaccactcccgccacccg atccaacccg 120 gtacttctcg acggaaaagt acatcgcact gagcaaagatgagaaattca aatttgatga 180 ttacgatgtg aatgatgaga cgctgaaaaa agtggtgctcaacgagattg gcaagtgccc 240 ggatatttgg agctcgcgga gccaggcagc cattatggagcactatccga ttgttgcaac 300 tgaaacgtac aggaggacag ggttgctgtt gtctatcaaatcgctgaaac aaatctacaa 360 atgcggaaag gacaatctcc gaaaccggct tcgcgtggcaattgtaagca agcggcttac 420 accggcccaa gtagaggcct atatgtggcg ctgggagttttacggcttta ttcgctacta 480 tcgagactat acacaacgct gggaggccga cttgttgaaagatttggacg tggtgctcgg 540 gctcgaggct cggcgagcat cgaaaaatat ggaaaaggtggattctgggg agctcatgga 600 gcccatggag cccatggatt ctacaatgga tgagatgtgcgtcgaggagg agccctacga 660 ggagacaggg tccaattgga gcgatccggc gccggaaccatcccaatcca aatcccagtc 720 cccagaagcc aagtaccctc aagcctacct actacctgaggcggacgaag tctacaatcc 780 tgacgatttc tatcaagagg aacatgaatc cgcatcaaacgccatgtatc ggatcgcttt 840 ctcacagcag tacggtggcg gcgggtcccc agccgtgcagaagcccgtca cttttagtgc 900 tcagccggcg ccggcgccag ttagagaggc cccaagcccagttgtggaga atgttagttc 960 atcgagtttc accccgaagc ccccggccat gatcaacaattttggtgagg agatgaacca 1020 aataacatac caagcgatcc gtattgcccg aaagcagccggaacgtctga aattgctccg 1080 taaggcactt ttcgacgttg tcctggcgtt tgatcagaaggaatacgccg atgttgggga 1140 tttgtacagg gatttggcgc aaaagaattc gtgataatttttttttgagt tttttaattt 1200 ttaatttatt tcaatttttg ttacatgttc caatataataaacaggtgct tgtttaaaaa 1260 aaaaaaaaaa aaaaaa 1276 47 386 PRTCaenorhabditis elegans 47 Met Ser Lys Ile Lys Thr His Ser Thr Gly SerLys Arg Thr Val Pro 1 5 10 15 Phe Tyr Lys Leu Pro Pro Pro Val Pro LeuPro Pro Leu Pro Pro Pro 20 25 30 Asp Pro Thr Arg Tyr Phe Ser Thr Glu LysTyr Ile Ala Leu Ser Lys 35 40 45 Asp Glu Lys Phe Lys Phe Asp Asp Tyr AspVal Asn Asp Glu Thr Leu 50 55 60 Lys Lys Val Val Leu Asn Glu Ile Gly LysCys Pro Asp Ile Trp Ser 65 70 75 80 Ser Arg Ser Gln Ala Ala Ile Met GluHis Tyr Pro Ile Val Ala Thr 85 90 95 Glu Thr Tyr Arg Arg Thr Gly Leu LeuLeu Ser Ile Lys Ser Leu Lys 100 105 110 Gln Ile Tyr Lys Cys Gly Lys AspAsn Leu Arg Asn Arg Leu Arg Val 115 120 125 Ala Ile Val Ser Lys Arg LeuThr Pro Ala Gln Val Glu Ala Tyr Met 130 135 140 Trp Arg Trp Glu Phe TyrGly Phe Ile Arg Tyr Tyr Arg Asp Tyr Thr 145 150 155 160 Gln Arg Trp GluAla Asp Leu Leu Lys Asp Leu Asp Val Val Leu Gly 165 170 175 Leu Glu AlaArg Arg Ala Ser Lys Asn Met Glu Lys Val Asp Ser Gly 180 185 190 Glu LeuMet Glu Pro Met Glu Pro Met Asp Ser Thr Met Asp Glu Met 195 200 205 CysVal Glu Glu Glu Pro Tyr Glu Glu Thr Gly Ser Asn Trp Ser Asp 210 215 220Pro Ala Pro Glu Pro Ser Gln Ser Lys Ser Gln Ser Pro Glu Ala Lys 225 230235 240 Tyr Pro Gln Ala Tyr Leu Leu Pro Glu Ala Asp Glu Val Tyr Asn Pro245 250 255 Asp Asp Phe Tyr Gln Glu Glu His Glu Ser Ala Ser Asn Ala MetTyr 260 265 270 Arg Ile Ala Phe Ser Gln Gln Tyr Gly Gly Gly Gly Ser ProAla Val 275 280 285 Gln Lys Pro Val Thr Phe Ser Ala Gln Pro Ala Pro AlaPro Val Arg 290 295 300 Glu Ala Pro Ser Pro Val Val Glu Asn Val Ser SerSer Ser Phe Thr 305 310 315 320 Pro Lys Pro Pro Ala Met Ile Asn Asn PheGly Glu Glu Met Asn Gln 325 330 335 Ile Thr Tyr Gln Ala Ile Arg Ile AlaArg Lys Gln Pro Glu Arg Leu 340 345 350 Lys Leu Leu Arg Lys Ala Leu PheAsp Val Val Leu Ala Phe Asp Gln 355 360 365 Lys Glu Tyr Ala Asp Val GlyAsp Leu Tyr Arg Asp Leu Ala Gln Lys 370 375 380 Asn Ser 385 48 1108 DNACaenorhabditis elegans 48 gcaaaaaact agatattttg tggcattttt acaattaaaaaacctttaaa aaatggatca 60 ccatgctatg taccgaaccg ctgaattcaa caaaactactgtccgattat tggcggaatt 120 catcgaaaag actgggcaga atgcgacgat agtgaatatggacagctttt tggagttctt 180 tgcgtatttg aatcccacgg ctccaattcc aacggttccagaaattgaaa aataattatt 240 gctaaaatca ccgattcgtt gcattgtgtg tggaatggaaactgaatcag attccgcagt 300 gacattaagc atcgataatg cttcaattat tctcacagcgacagtaattg gttactgtag 360 agatccaagt gatgcagtta atcaaattcg aaaggagagtcttcgagcat gcacgaaaca 420 tttcaacagt attttccatg tcatcttcga aggactgcaaatcgagaaca cctactgtgc 480 tcatcatgca aaatacagtc ttgccaatcg ttggtgcaaagtctacacga tgattcgatc 540 ttccctgggc gagcagttca caaagttcga tgtgcgcaattttaaatcaa tattgcaatc 600 atttttggat acttttggtg aaattgatga cgacaaaaaggataaagaat cttctcattt 660 tgatgaatgt tttgaagaaa tggattcaga aaacgtagaaattaaaatgg agagcccaca 720 agaagaagct gcagagaaat cgaagttttc tgaaaacctagtggaggtaa aactggaacc 780 aattgaaact catgaacttg acaaaactat atccgacttttcttcaagtg atataattga 840 ttcgtcccaa aaactgcagc aaaatggttt tcctgaaaaagtggagcaaa tggacaaata 900 tagcaacaaa ttgaaagatg aagcttcaga caaaaagtatgaaaagccag gaaaaaagga 960 ctacgttgaa gaagagggat actgggcgcc gatcaccgacagcgaggatg atgaagcctg 1020 aatttattta atcaaacgtt ttggaaattt tttttgtttttgtcaataaa accatataac 1080 aataaaaaaa aaaaaaaaaa aactcgag 1108 49 60 PRTCaenorhabditis elegans 49 Met Asp His His Ala Met Tyr Arg Thr Ala GluPhe Asn Lys Thr Thr 1 5 10 15 Val Arg Leu Leu Ala Glu Phe Ile Glu LysThr Gly Gln Asn Ala Thr 20 25 30 Ile Val Asn Met Asp Ser Phe Leu Glu PhePhe Ala Tyr Leu Asn Pro 35 40 45 Thr Ala Pro Ile Pro Thr Val Pro Glu IleGlu Lys 50 55 60 50 50 000 51 60 PRT Caenorhabditis elegans 51 Arg CysIle Val Cys Gly Met Glu Thr Glu Ser Asp Ser Ala Val Thr 1 5 10 15 LeuSer Ile Asp Asn Ala Ser Ile Ile Leu Thr Ala Thr Val Ile Gly 20 25 30 TyrCys Arg Asp Pro Ser Asp Ala Val Asn Gln Ile Arg Lys Glu Ser 35 40 45 LeuArg Ala Cys Thr Lys His Phe Asn Ser Ile Phe 50 55 60 52 60 PRTCaenorhabditis elegans 52 Pro Cys Ile Leu Cys Glu Lys Ala Leu Leu MetArg Glu Ser Ile Ala 1 5 10 15 Met Thr Asp Asn Glu Ala Val Lys Val LeuMet Ala Ala Val Met Ser 20 25 30 Gly His Phe Arg Met Ala Thr Ala Glu LysAla Ile Arg His Glu Arg 35 40 45 Leu Arg Met Cys Tyr Asp His Val Asp PheVal Tyr 50 55 60 53 60 PRT Caenorhabditis elegans 53 Pro Cys Ile Ile CysGly Asn Glu Val Pro Gly His Arg Ser Ile Arg 1 5 10 15 Val Ser Asp AspAsp Ala Ala Ile Phe Leu Thr Ala Ala Val Leu Thr 20 25 30 Asp Gln Lys ThrIle Arg Gln Ala Lys Arg Asp Ile Leu Ser Glu Tyr 35 40 45 Leu Thr Val CysLeu Arg His Ser Leu His Tyr Tyr 50 55 60 54 60 PRT Caenorhabditiselegans 54 Pro Cys Leu Val Cys Asn Gln Gln Met Glu Met Thr Lys Val ArgSer 1 5 10 15 Val Asn Asn Thr Asp Ala Tyr Ile Met Ile Tyr Val Cys ValMet Asn 20 25 30 Asp Lys Tyr Asp Met Asp Lys Ala Lys Glu Leu Ala Arg MetGln Arg 35 40 45 Phe Lys Cys Cys Val Ser His Leu Asp Glu Leu Tyr 50 5560 55 177 PRT Caenorhabditis elegans 55 Met Leu Ser Ile Lys Gln Glu LeuLeu Asp Ala Pro Pro Pro Pro Pro 1 5 10 15 Ala Ala Thr Pro Leu Pro ProIle Thr His Arg Ile Ser Leu Ser Gly 20 25 30 Tyr Arg Asn Ile His Ala LysSer Phe Leu Lys Thr Met Thr Met Asp 35 40 45 Leu Cys Val Arg Arg Val ValLeu Ser Leu Leu Glu Asn Arg Arg Ala 50 55 60 Leu Trp Ile Arg Val His LysSer Pro Lys Ala Asp Trp Glu Val Leu 65 70 75 80 Gly Val Glu Val Phe GluArg Thr Gly Lys Ala Val Ser Val Lys Gln 85 90 95 Leu Gln Arg Ile Phe LeuThr Ala Arg Asp Trp Leu Arg Arg Asn Leu 100 105 110 Gln Leu Tyr Ile IleGln Arg Lys Met Asp Lys Leu Thr Leu Asp Ala 115 120 125 Glu Leu Ala LysTrp Glu Leu Tyr Pro His Phe Ile Tyr Tyr Arg Gln 130 135 140 Tyr Leu GlyGln Phe Glu Ala His Leu Arg Gly Glu Glu Trp Thr Gly 145 150 155 160 GluLeu Tyr Asp Asp Asp Ile Ile Cys Asp Gly Ile Met Gln Val Glu 165 170 175Val 56 75 PRT Caenorhabditis elegans 56 Glu Asp Ser Val Ser Tyr Thr LysIle Thr Glu Asp Leu Leu Gln Lys 1 5 10 15 Lys Pro His Lys His Arg PheIle Arg Gln Ala Leu Phe Lys Thr Ile 20 25 30 Met Ala Leu Asp Asp Asp GluVal Glu Tyr Thr Glu Leu Ala Asp Leu 35 40 45 Phe Gly Asp Ile Ala Glu GlnSer Asn Val Val Arg Arg Leu Arg Leu 50 55 60 Gln Arg Gln Gln Gln Arg GlyArg Gly Glu Gln 65 70 75 57 178 PRT Caenorhabditis elegans 57 Met SerLeu Ile Lys Gln Glu His Met His Pro Pro Pro Arg Ala Ile 1 5 10 15 ThrPro Leu Pro Pro Ala Thr His Gln Ile Thr Leu Glu Glu Tyr Lys 20 25 30 GluArg Glu Lys Lys Asp Tyr Tyr Arg Asp Ala Thr Lys Asp Ala Ser 35 40 45 ValLys Lys Val Val Leu Ser Leu Leu Lys Asp His Pro Gly Met Trp 50 55 60 GlnAsn Gly Asn Arg Phe Gln Pro Glu Lys Trp Arg Ala Leu Gly Val 65 70 75 80Asp Val Tyr Gln Arg Thr Gly Gln Ile Val Arg Val Asn Asp Met Arg 85 90 95Lys Met Leu Val Met Gly Lys Ser Val Leu Lys Lys Lys Ile Ala Ile 100 105110 Cys Ile Arg Asp Lys Lys Leu Asp Arg Ala Ala Thr Glu Lys Asp Leu 115120 125 Trp Tyr Trp Glu Tyr Tyr Arg His Phe Leu Tyr Tyr Arg Glu Thr Leu130 135 140 Gly Gln Phe Glu Ala Asn Leu Arg Gly Glu Glu Trp Thr Gly GluAsp 145 150 155 160 Gln Ile Gln Asp Glu Asp Asp Ile Ile Tyr Asp Gly MetLeu Asp Gly 165 170 175 Asp Leu 58 73 PRT Caenorhabditis elegans 58 ArgSer Ala Gln His Ile Ala Glu Gln Ala Lys Arg Leu Phe Leu Gln 1 5 10 15Tyr Pro Glu Lys Ser Asn Leu Ile Arg Glu Thr Met Phe Lys Thr Ile 20 25 30Leu Ala Phe Asp Asp Pro Ser Ala Asp Tyr Gln Asn Val Gly Glu Ile 35 40 45Phe Asp Asp Leu Ala Ala Gln Glu Ala Ala Lys Lys Arg Lys Arg Ala 50 55 60Glu Asn Arg Ala Gln Arg Glu Gln Gln 65 70 59 179 PRT Caenorhabditiselegans 59 Met Ser Leu Ile Lys Gln Glu His Met Asn Pro Pro Pro Arg ThrIle 1 5 10 15 Thr Pro Leu Pro Pro Pro Thr His Gln Ile Thr Ile Glu GluTyr Lys 20 25 30 Glu Arg Val Lys Arg Asp Tyr Tyr Arg Asn Ala Thr Lys AspThr Ser 35 40 45 Leu Lys Lys Val Val Leu Ser Leu Ile Lys Asp Arg Lys AlaMet Trp 50 55 60 Ala Pro Ala Ala Lys Pro Ser Glu Asp Lys Trp Gln Lys LeuGly Ala 65 70 75 80 Glu Val Phe Ser Arg Thr Gly Lys Val Val Ser Val ThrGln Leu Arg 85 90 95 Arg Met Leu Val Ser Ser Lys His Val Leu Lys Thr LysMet Ser His 100 105 110 Cys Ile Lys Val Lys Lys Met Asp Arg Val Ser ThrGlu Ala Tyr Leu 115 120 125 Trp Asn Trp Glu Phe Tyr Arg His Phe Leu TyrTyr Arg Glu Met Leu 130 135 140 Asp Arg Phe Glu Ala Asn Leu Arg Gly LysGln Trp Thr Gly Glu Asp 145 150 155 160 Gln Pro Thr Asp Asp Asp Asp AspIle Ile Cys Asp Gly Ile Phe Glu 165 170 175 Val Glu Met 60 70 PRTCaenorhabditis elegans 60 Ser Thr Ala Glu Gln Ile Gly Glu Glu Ile AspArg Leu Ile Gln Leu 1 5 10 15 Tyr Pro Gln Arg Glu Met Leu Ile Arg GlnAla Phe Phe Lys Thr Ile 20 25 30 Phe Ala Leu Glu Asp Glu Thr Val Glu PheSer Asn Leu Gly Asp Leu 35 40 45 Phe Glu Asp Leu Ala Glu Gln Glu Asn PheLys Arg Arg Arg Arg Ser 50 55 60 Arg Ala Gln Arg Leu Glu 65 70 61 187PRT Caenorhabditis elegans 61 Met Leu Asn Ile Lys Gln Glu Gly Val ValAla Asp Ala Pro Arg Ala 1 5 10 15 Leu Thr Pro Ile Pro Pro Phe Ile HisHis Val Ser Met Glu Glu Tyr 20 25 30 Met Gly Met Glu Leu Asn Ser Val TyrGlu Glu Ala Thr Lys Asp Ser 35 40 45 Ala Leu Lys Lys Val Val Leu Asp LeuLeu Lys Asp Arg Pro Gly Met 50 55 60 Trp Gln Asn Gly Asn Arg Phe Gln LeuGlu Asn Trp Arg Glu Leu Gly 65 70 75 80 Val Asp Val Tyr Gln Arg Thr GlyGln Ile Val Arg Ala Glu Leu Gly 85 90 95 Glu Val Ser Val Asn Asp Met HisArg Met Phe Val Val Gly Lys Ala 100 105 110 Val Leu Lys Gln Lys Ile ThrVal Cys Ile Arg Tyr Lys Lys Leu Asp 115 120 125 Arg Ala Ala Thr Glu AlaAsp Leu Gln Asn Trp Glu Phe Tyr Arg His 130 135 140 Phe Arg Tyr Tyr ArgGlu Thr Leu Gly Gln Phe Glu Ala Asn Leu Arg 145 150 155 160 Gly Glu GlnTrp Thr Gly Glu Asp Gln Pro Ala Asp Asp Asp Asp Asp 165 170 175 Ile IleTyr Asp Gly Ile Phe Glu Val Glu Met 180 185 62 69 PRT Caenorhabditiselegans 62 Ser Thr Ala Glu Gln Ile Gly Glu Glu Ile Asp Arg Leu Ile GlnLeu 1 5 10 15 Tyr Pro Gln Arg Glu Met Leu Ile Arg Gln Ala Phe Phe LysThr Ile 20 25 30 Phe Ala Leu Glu Asp Glu Thr Val Glu Phe Ser Asn Leu GlyAsp Leu 35 40 45 Phe Glu Asp Leu Ala Glu Gln Glu Asn Phe Lys Arg Arg ArgArg Ser 50 55 60 Ala Gln Arg Leu Glu 65 63 186 PRT Caenorhabditiselegans 63 Met Met Asn Pro Lys Glu Glu Pro Arg Pro Phe Ser Ile Val ProLeu 1 5 10 15 Pro Arg Pro Pro Arg Pro Thr Thr Pro Leu Pro Pro Ile SerHis Cys 20 25 30 Ile Thr Met Ala Asp Tyr Leu Leu Leu Glu Asn Thr Lys PheHis Lys 35 40 45 Thr Ala Thr Arg Ala Pro Lys Ile Lys Lys Val Leu Leu SerLeu Leu 50 55 60 Lys Asp Arg Pro Glu Ile Trp Asp Arg Lys Ala Gln Phe SerAla Lys 65 70 75 80 Asn Trp Gln Asn Leu Gly Val Glu Val Tyr Glu Arg ThrGly Tyr Ile 85 90 95 Val Arg Ser Asn Asp Leu His Lys Met Leu Arg Thr AlaLys Val Val 100 105 110 Leu Lys Asn Lys Leu Arg Thr Cys Ile Gly Ile LysLys Leu Asp Arg 115 120 125 Ala Ala Thr Glu Thr Glu Leu Trp Lys Trp GluTyr Tyr Pro His Phe 130 135 140 Ile Tyr Tyr Arg Glu Thr Leu Gly His PheGlu Ala Asn Leu Arg Gly 145 150 155 160 Glu Pro Trp Asp Gly Glu Ala HisIle Asp Asp Asp Asp Asp Asp Ile 165 170 175 Ile Tyr Glu Gly Tyr Trp GluAla Asp Lys 180 185 64 70 PRT Caenorhabditis elegans 64 Asn Ser Ala GlnHis Ile Gly Glu Gln Val His Arg Leu Phe Ala Gln 1 5 10 15 Tyr Pro GluArg Ser Lys Leu Phe Arg Glu Thr Leu Phe Lys Thr Ile 20 25 30 Leu Ala LeuGlu Glu Pro Glu Tyr Glu His Ala Ala Glu Val Phe Thr 35 40 45 Asp Leu AlaGln Ser Glu Thr Ala Lys Arg Arg Arg Arg Ser Glu Ala 50 55 60 Thr Trp GlnAsn Gly Gln 65 70 65 186 PRT Caenorhabditis elegans 65 Met Val Ser AlaThr Arg Val Pro Arg Arg Ser Ser Thr Thr Thr Ser 1 5 10 15 Ala Thr AlaGln Gln Arg Thr Pro Ser Pro Leu Met Pro Ala Ser Phe 20 25 30 Pro Ile ThrMet Asp Glu Tyr Leu Glu Lys Glu Asn Arg Glu Phe Val 35 40 45 Val Asn AlaSer Lys Asp Ile Ala Met Lys Lys Leu Ala Leu Thr Leu 50 55 60 Leu Glu LeuTyr Pro Glu Met Trp Lys Pro Gly Gly Pro Met Val Ala 65 70 75 80 Lys LysTrp Gln Ala Phe Gly Ala Glu Met Tyr Arg Arg Thr Gly Lys 85 90 95 Ile TyrArg Cys Lys Asp Leu His Ser Val Phe Thr Leu Thr Lys Ser 100 105 110 SerIle Lys Arg Lys Leu Arg Thr Cys Ile Leu Ile Lys Arg Met His 115 120 125Arg Ser Lys Thr Asp Glu Glu Met Trp Lys Tyr Glu Leu Tyr Pro Tyr 130 135140 Phe Gln Tyr Tyr Arg Gln Ser Ile Gly Gln Phe Glu Ala Lys Leu Arg 145150 155 160 Asp Glu Pro Trp Thr Gly Glu Asp Gln Ala Gln Glu Asp Asp AspIle 165 170 175 Leu Phe Asp Gly Leu Phe Glu Val Glu Asn 180 185 66 66PRT Caenorhabditis elegans 66 Lys Thr Ala Asp Asn Ile Gly Asp Gln ValLys Gln Leu Phe Val Asp 1 5 10 15 His Pro Asp Arg Ala Asn Phe Phe ArgGlu Val Leu Phe Lys Thr Val 20 25 30 Leu Glu Leu Arg Asp Pro Ala Phe ThrAsn Ala Gly Val Phe Phe Asp 35 40 45 Glu Met Ser Ser Leu Glu Ser Ala LysArg Arg Arg Arg Ser Glu Met 50 55 60 Asn Lys 65 67 178 PRTCaenorhabditis elegans 67 Met Ser Arg Ile Lys Gln Glu Gln Val Asn ProPro Pro Pro Pro Arg 1 5 10 15 Ala Ile Thr Pro Leu Pro Pro Ala Thr HisArg Ile Thr Met Asp Glu 20 25 30 Tyr Lys Lys Arg Glu Lys Lys Asp Tyr TyrArg Asp Ala Thr Lys Asp 35 40 45 Ala Ser Val Lys Lys Val Val Leu Ser LeuLeu Lys Asp Tyr Pro Asp 50 55 60 Met Trp Gln Asn Gly Asn Arg Phe Gln ThrArg Lys Trp Arg Ala Leu 65 70 75 80 Gly Val Glu Val Tyr Gln Arg Thr GlyGln Ile Val Gly Val Asp Asp 85 90 95 Met Arg Lys Met Phe Met Ser Gly LysThr Val Leu Lys Gln Lys Ile 100 105 110 Thr Phe Cys Ile Arg Asn Met LysMet Asp Arg Ala Ala Thr Glu Ala 115 120 125 Asp Leu Gln Asn Trp Glu TyrTyr Arg His Phe Leu Tyr Tyr Arg Gln 130 135 140 Thr Leu Gly Lys Phe GluAla Lys Leu Arg Gly Glu Gln Trp Ile Gly 145 150 155 160 Glu Asp Gln ValGlu Asp Asp Asp Glu Asp Asp Val Ile Phe Asp Gly 165 170 175 Glu Ser 68410 PRT Caenorhabditis elegans 68 Met Gly Thr Cys Trp Gly Asp Ile SerGlu Asn Val Arg Val Glu Val 1 5 10 15 Pro Asn Thr Asp Cys Ser Leu ProThr Lys Val Phe Trp Ile Ala Gly 20 25 30 Ile Val Lys Leu Ala Gly Tyr AsnAla Leu Leu Arg Tyr Glu Gly Phe 35 40 45 Glu Asn Asp Ser Gly Leu Asp PheTrp Cys Asn Ile Cys Gly Ser Asp 50 55 60 Ile His Pro Val Gly Trp Cys AlaAla Ser Gly Lys Pro Leu Val Pro 65 70 75 80 Pro Arg Thr Ile Gln His LysTyr Thr Asn Trp Lys Ala Phe Leu Val 85 90 95 Lys Arg Leu Thr Gly Ala LysThr Leu Pro Pro Asp Phe Ser Gln Lys 100 105 110 Val Ser Glu Ser Met GlnTyr Pro Phe Lys Pro Cys Met Arg Val Glu 115 120 125 Val Val Asp Lys ArgHis Leu Cys Arg Thr Arg Val Ala Val Val Glu 130 135 140 Ser Val Ile GlyGly Arg Leu Arg Leu Val Tyr Glu Glu Ser Glu Asp 145 150 155 160 Arg ThrAsp Asp Phe Trp Cys His Met His Ser Pro Leu Ile His His 165 170 175 IleGly Trp Ser Arg Ser Ile Gly His Arg Phe Lys Arg Ser Asp Ile 180 185 190Thr Lys Lys Gln Asp Gly His Phe Thr Asp Pro Pro His Leu Phe Ala 195 200205 Lys Val Lys Glu Val Asp Gln Ser Gly Glu Trp Phe Lys Glu Gly Met 210215 220 Lys Leu Glu Ala Ile Asp Pro Leu Asn Leu Ser Thr Ile Cys Val Ala225 230 235 240 Thr Ile Lys Arg Val Leu Ala Asp Gly Phe Leu Met Ile GlyIle Asp 245 250 255 Gly Ser Glu Ala Ala Asp Gly Ser Asp Trp Phe Cys TyrHis Ala Thr 260 265 270 Ser Pro Ser Ile Phe Pro Val Gly Phe Cys Glu IleAsn Met Ile Glu 275 280 285 Leu Thr Pro Pro Arg Gly Tyr Thr Lys Leu ProPhe Lys Trp Phe Asp 290 295 300 Tyr Leu Arg Glu Thr Gly Ser Ile Ala AlaPro Val Lys Leu Phe Asn 305 310 315 320 Lys Asp Val Pro Asn His Gly PheArg Val Gly Met Lys Leu Glu Ala 325 330 335 Val Asp Leu Met Glu Pro ArgLeu Ile Cys Val Ala Thr Val Thr Arg 340 345 350 Ile Ile His Arg Leu LeuArg Ile His Phe Asp Gly Trp Glu Glu Glu 355 360 365 Tyr Asp Gln Trp ValAsp Cys Glu Ser Pro Asp Leu Tyr Pro Val Gly 370 375 380 Trp Cys Gln LeuThr Gly Tyr Gln Leu Gln Pro Pro Ala Ser Gln Cys 385 390 395 400 Lys LeuVal Tyr Arg Lys Gly Val Leu Leu 405 410 69 512 PRT Caenorhabditiselegans 69 Met Asn Phe Ser Asn Lys Lys Val Ile Leu Lys Ala Phe Leu SerLys 1 5 10 15 Asn Ile Ile Tyr Tyr Phe Gln Arg Gln Tyr Asn Tyr Lys LeuGlu Glu 20 25 30 Ala Glu Tyr Arg Tyr Phe Thr Glu Glu Arg Leu Phe Tyr ArgArg Arg 35 40 45 Asn Pro Val Glu Lys Ile Ala Gln Arg Ile Pro Lys Pro GlnIle Glu 50 55 60 Gly Thr Phe Thr Trp Ser Asp Glu Leu Arg Cys Asn Tyr AspGly Asn 65 70 75 80 Thr Gln Phe Leu Pro Val Glu Ala Leu Glu Gly Cys LeuPro Leu Glu 85 90 95 Lys Leu Asn Gln His Leu Lys Pro Gly Phe Arg Leu GluVal Val Val 100 105 110 Arg Pro Ser Leu Asp Pro Ser Ile Thr Thr Lys SerPro Glu Ile Arg 115 120 125 Trp Phe Gly Glu Val Thr Ala Val Cys Gly PheTyr Val Ala Ile Lys 130 135 140 Phe Val Gly Glu Leu Asn Arg Arg Pro CysTrp Phe His Met Leu Ser 145 150 155 160 Glu Asp Ile Phe Asp Ile Gly SerGly Leu Lys Gln Asp Pro Ala Met 165 170 175 Lys Trp Leu Gln Tyr Arg ProLeu Ser Leu Leu Lys Pro Met Gln Cys 180 185 190 Pro Lys Phe Trp Arg ArgGly Ser Thr Pro Ala Pro Pro Val Pro Arg 195 200 205 Pro Thr Glu Glu IleLeu Asp Glu Phe Gln Ala Glu Leu His Glu Asn 210 215 220 Arg Ile Ser GluPro Lys Ile Phe Asp Gln Leu Arg His Leu Ala His 225 230 235 240 Arg ProSer Arg Phe Arg Leu Asn Gln Arg Val Glu Leu Leu Asn Tyr 245 250 255 LeuGlu Pro Thr Glu Ile Arg Val Ala Arg Ile Leu Arg Ile Leu Gly 260 265 270Arg Arg Leu Met Val Met Val Thr Ala Gln Asp Tyr Pro Glu Asp Leu 275 280285 Pro Ser Val Glu Ala Lys Asp Arg Gln Val Gln His Glu Asn Val Glu 290295 300 Phe Trp Val Asp Glu Ser Ser Phe Phe Leu Phe Pro Val Gly Phe Ala305 310 315 320 Met Ile Asn Gly Leu Arg Thr Lys Ala Thr Glu Gly Tyr LeuGlu His 325 330 335 Ser Arg Arg Ile Ala Glu Gly Ser Gly Thr Glu Lys LeuAsn Leu Leu 340 345 350 Lys Val Gly Gln Lys Phe Glu Leu Leu Asp Pro LeuSer Asp Leu Arg 355 360 365 Gln Ser Phe Cys Val Ala Thr Ile Arg Lys IleCys Lys Thr Pro Gly 370 375 380 Phe Leu Ile Ile Ser Pro Asp Glu Thr GluSer Asp Asp Glu Ser Phe 385 390 395 400 Pro Ile His Ile Asp Asn His PheMet His Pro Val Gly Tyr Ala Glu 405 410 415 Lys Phe Gly Ile Lys Leu AspArg Leu Ala Gly Thr Glu Pro Gly Lys 420 425 430 Phe Lys Trp Glu Gly TyrLeu Lys Glu Lys Gln Ala Glu Lys Ile Pro 435 440 445 Asp Glu Met Leu ArgPro Leu Pro Ser Lys Glu Arg Arg His Met Phe 450 455 460 Glu Phe Gly ArgVal Leu Glu Ala Val Gly Gln Asn Glu Thr Tyr Trp 465 470 475 480 Ile SerPro Ala Ser Val Glu Glu Val His Gly Arg Thr Val Leu Ile 485 490 495 GluPhe Gln Gly Trp Asp Ser Glu Phe Ser Glu Leu Tyr Asp Met Glu 500 505 51070 411 PRT Caenorhabditis elegans 70 Met Ser Glu Phe Leu Lys Ile Val ArgAla Asn Lys Lys Ser Asp Arg 1 5 10 15 Lys Leu Asp Lys Thr Tyr Leu TrpGlu Ser Tyr Leu His Gln Phe Glu 20 25 30 Lys Gly Lys Thr Ser Phe Ile ProVal Glu Ala Phe Asn Arg Asn Leu 35 40 45 Thr Val Asn Phe Asn Glu Cys ValLys Glu Gly Val Ile Phe Glu Thr 50 55 60 Val Val His Asp Tyr Asp Lys AsnCys Asp Ser Ile Gln Val Arg Trp 65 70 75 80 Phe Ala Arg Ile Glu Lys ValCys Gly Tyr Arg Val Leu Ala Gln Phe 85 90 95 Ile Gly Ala Asp Thr Lys PheTrp Leu Asn Ile Leu Ser Asp Asp Met 100 105 110 Phe Gly Leu Ala Asn AlaAla Met Ser Asp Pro Asn Met Asp Lys Ile 115 120 125 Val Tyr Ala Pro ProLeu Ala Ile Asn Glu Glu Tyr Gln Asn Asp Met 130 135 140 Val Asn Tyr ValAsn Asn Cys Ile Asp Gly Glu Ile Val Gly Gln Thr 145 150 155 160 Ser LeuSer Pro Lys Phe Asp Glu Gly Lys Ala Leu Leu Ser Lys His 165 170 175 ArgPhe Lys Val Gly Gln Arg Leu Glu Leu Leu Asn Tyr Ser Asn Ser 180 185 190Thr Glu Ile Arg Val Ala Arg Ile Gln Glu Ile Cys Gly Arg Arg Met 195 200205 Asn Val Ser Ile Thr Lys Lys Asp Phe Pro Glu Ser Leu Pro Asp Ala 210215 220 Asp Asp Asp Arg Gln Val Phe Ser Ser Gly Ser Gln Tyr Trp Ile Asp225 230 235 240 Glu Gly Ser Phe Phe Ile Phe Pro Val Gly Phe Ala Ala ValAsn Gly 245 250 255 Tyr Gln Leu Asn Ala Lys Lys Glu Tyr Ile Glu His ThrAsn Lys Ile 260 265 270 Ala Gln Ala Ile Lys Asn Gly Glu Asn Pro Arg TyrAsp Ser Asp Asp 275 280 285 Val Thr Phe Asp Gln Leu Ala Lys Asp Pro IleAsp Pro Met Ile Trp 290 295 300 Arg Lys Val Lys Val Gly Gln Lys Phe GluLeu Ile Asp Pro Leu Ala 305 310 315 320 Gln Gln Phe Asn Asn Leu His ValAla Ser Ile Leu Lys Phe Cys Lys 325 330 335 Thr Glu Gly Tyr Leu Ile ValGly Met Asp Gly Pro Asp Ala Leu Glu 340 345 350 Asp Ser Phe Pro Ile HisIle Asn Asn Thr Phe Met Phe Pro Val Gly 355 360 365 Tyr Ala Glu Lys TyrAsn Leu Glu Leu Val Pro Pro Asp Glu Phe Lys 370 375 380 Gly Thr Phe ArgTrp Asp Glu Tyr Leu Glu Lys Glu Ser Ala Glu Thr 385 390 395 400 Leu ProLeu Asp Leu Phe Lys Pro Met Pro Ser 405 410 71 498 PRT Caenorhabditiselegans 71 Met Ser Glu Phe Leu Lys Ile Val Arg Ala Asn Lys Lys Ser AspArg 1 5 10 15 Lys Leu Asp Lys Thr Tyr Leu Trp Glu Ser Tyr Leu His GlnPhe Glu 20 25 30 Lys Gly Lys Thr Ser Phe Ile Pro Val Glu Ala Phe Asn ArgAsn Leu 35 40 45 Thr Val Asn Phe Asn Glu Cys Val Lys Glu Gly Val Ile PheGlu Thr 50 55 60 Val Val His Asp Tyr Asp Lys Asn Cys Asp Ser Ile Gln ValArg Trp 65 70 75 80 Phe Ala Arg Ile Glu Lys Val Cys Gly Tyr Arg Val LeuAla Gln Phe 85 90 95 Ile Gly Ala Asp Thr Lys Phe Trp Leu Asn Ile Leu SerAsp Asp Met 100 105 110 Phe Gly Leu Ala Asn Ala Ala Met Ser Asp Pro AsnMet Asp Lys Ile 115 120 125 Val Tyr Ala Pro Pro Leu Ala Ile Asn Glu GluTyr Gln Asn Asp Met 130 135 140 Val Asn Tyr Val Asn Asn Cys Ile Asp GlyGlu Ile Val Gly Gln Thr 145 150 155 160 Ser Leu Ser Pro Lys Phe Asp GluGly Lys Ala Leu Leu Ser Lys His 165 170 175 Arg Phe Lys Val Gly Gln ArgLeu Glu Leu Leu Asn Tyr Ser Asn Ser 180 185 190 Thr Glu Ile Arg Val AlaArg Ile Gln Glu Ile Cys Gly Arg Arg Met 195 200 205 Asn Val Ser Ile ThrLys Lys Asp Phe Pro Glu Ser Leu Pro Asp Ala 210 215 220 Asp Asp Asp ArgGln Val Phe Ser Ser Gly Ser Gln Tyr Trp Ile Asp 225 230 235 240 Glu GlySer Phe Phe Ile Phe Pro Val Gly Phe Ala Ala Val Asn Gly 245 250 255 TyrGln Leu Asn Ala Lys Lys Glu Tyr Ile Glu His Thr Asn Lys Ile 260 265 270Ala Gln Ala Ile Lys Asn Gly Glu Asn Pro Arg Tyr Asp Ser Asp Asp 275 280285 Val Thr Phe Asp Gln Leu Ala Lys Asp Pro Ile Asp Pro Met Ile Trp 290295 300 Arg Lys Val Lys Val Gly Gln Lys Phe Glu Leu Ile Asp Pro Leu Ala305 310 315 320 Gln Gln Phe Asn Asn Leu His Val Ala Ser Ile Leu Lys PheCys Lys 325 330 335 Thr Glu Gly Tyr Leu Ile Val Gly Met Asp Gly Pro AspAla Leu Glu 340 345 350 Asp Asn Phe Pro Ile His Ile Asn Asn Thr Phe MetPhe Pro Val Gly 355 360 365 Tyr Ala Glu Lys Tyr Asn Leu Glu Leu Val ProPro Asp Glu Phe Lys 370 375 380 Gly Thr Phe Arg Trp Asp Glu Tyr Leu GluLys Glu Ser Ala Glu Thr 385 390 395 400 Leu Pro Leu Asp Leu Phe Lys ProMet Pro Ser Gln Glu Arg Leu Asp 405 410 415 Lys Phe Lys Val Ile Leu IleSer Lys Arg Val Gly Leu Arg Leu Glu 420 425 430 Ala Ala Asp Met Cys GluAsn Gln Phe Ile Cys Pro Ala Thr Val Lys 435 440 445 Ser Val His Gly ArgLeu Ile Asn Val Asn Phe Asp Gly Trp Asp Glu 450 455 460 Glu Phe Asp GluLeu Tyr Asp Val Asp Ser His Asp Ile Leu Pro Ile 465 470 475 480 Gly TrpCys Glu Ala His Ser Tyr Val Leu Gln Pro Pro Lys Lys Tyr 485 490 495 AsnTyr 72 498 PRT Caenorhabditis elegans 72 Met Ser Glu Phe Leu Lys Ile ValArg Ala Asn Lys Lys Ser Asp Arg 1 5 10 15 Lys Leu Asp Lys Thr Tyr LeuTrp Glu Ser Tyr Leu His Gln Phe Glu 20 25 30 Lys Gly Lys Thr Ser Phe IlePro Val Glu Ala Phe Asn Arg Asn Leu 35 40 45 Thr Val Asn Phe Asn Glu CysVal Lys Glu Gly Val Ile Phe Glu Thr 50 55 60 Val Val His Asp Tyr Asp LysAsn Cys Asp Ser Ile Gln Val Arg Trp 65 70 75 80 Phe Ala Arg Ile Glu LysVal Cys Gly Tyr Arg Val Leu Ala Gln Phe 85 90 95 Ile Gly Ala Asp Thr LysPhe Trp Leu Asn Ile Leu Ser Asp Asp Met 100 105 110 Phe Gly Leu Ala AsnAla Ala Met Ser Asp Pro Asn Met Asp Lys Ile 115 120 125 Val Tyr Ala SerPro Leu Ala Ile Asn Glu Glu Tyr Gln Asn Asp Met 130 135 140 Val Asn TyrVal Asn Asn Cys Ile Asp Gly Glu Ile Val Gly Gln Thr 145 150 155 160 SerLeu Ser Pro Lys Phe Asp Glu Gly Lys Ala Leu Leu Ser Lys His 165 170 175Arg Phe Lys Val Gly Gln Arg Leu Glu Leu Leu Asn Tyr Ser Asn Ser 180 185190 Thr Glu Ile Arg Val Ala Arg Ile Gln Glu Ile Cys Gly Arg Arg Met 195200 205 Asn Val Ser Ile Thr Lys Lys Asp Phe Pro Glu Ser Leu Pro Asp Ala210 215 220 Asp Asp Asp Arg Gln Val Phe Ser Ser Gly Ser Gln Tyr Trp IleAsp 225 230 235 240 Glu Gly Ser Phe Phe Ile Phe Pro Val Gly Phe Ala AlaVal Asn Gly 245 250 255 Tyr Gln Leu Asn Ala Lys Lys Glu Tyr Ile Glu HisThr Asn Lys Ile 260 265 270 Ala Gln Ala Ile Lys Asn Gly Glu Asn Pro ArgTyr Asp Ser Asp Asp 275 280 285 Val Thr Phe Asp Gln Leu Ala Lys Asp ProIle Asp Pro Met Ile Trp 290 295 300 Arg Lys Val Lys Val Gly Gln Lys PheGlu Leu Ile Asp Pro Leu Ala 305 310 315 320 Gln Gln Phe Asn Asn Leu HisVal Ala Ser Ile Leu Lys Phe Cys Lys 325 330 335 Thr Glu Gly Tyr Leu IleVal Gly Met Asp Gly Pro Asp Ala Leu Glu 340 345 350 Asp Ser Phe Pro IleHis Ile Asn Asn Thr Phe Met Phe Pro Val Gly 355 360 365 Tyr Ala Glu LysTyr Asn Leu Glu Leu Val Pro Pro Asp Glu Phe Lys 370 375 380 Gly Thr PheArg Trp Asp Glu Tyr Leu Glu Lys Glu Ser Ala Glu Thr 385 390 395 400 LeuPro Leu Asp Leu Phe Lys Pro Met Pro Ser Gln Glu Arg Leu Asp 405 410 415Lys Phe Lys Val Ile Leu Ile Ser Lys Arg Val Gly Leu Arg Leu Glu 420 425430 Ala Ala Asp Met Cys Glu Asn Gln Phe Ile Cys Pro Ala Thr Val Lys 435440 445 Ser Val His Gly Arg Leu Ile Asn Val Asn Phe Asp Gly Trp Asp Glu450 455 460 Glu Phe Asp Glu Leu Tyr Asp Val Asp Ser His Asp Ile Leu ProIle 465 470 475 480 Gly Trp Cys Glu Ala His Ser Tyr Val Leu Gln Pro ProLys Lys Tyr 485 490 495 Asn Tyr 73 1497 DNA Caenorhabditis elegans 73atgtctgaat ttctgaaaat tgtcagagct aacaaaaaat cggacagaaa actcgataag 60acctacttgt gggaatccta tttacatcag ttcgagaaag gaaaaacttc tttcattcca 120gttgaagcat tcaatcgtaa ccttacagtt aattttaacg aatgcgtgaa ggaaggagtt 180atcttcgaaa cagtggtcca tgattatgac aagaactgcg attcgattca agtcagatgg 240tttgcacgaa ttgaaaaagt ttgcggatac agagttctgg ctcagtttat cggagctgac 300acgaaatttt ggctcaatat tttatcggac gatatgtttg gtttggcaaa cgccgcaatg 360agtgatccca atatggataa aattgtatat gctccgccgc ttgcaatcaa cgaagaatac 420caaaatgata tggtaaatta tgtaaataat tgcattgatg gcgaaatcgt cggccaaact 480tcgctgtctc caaaattcga tgaagggaag gctctcctaa gcaagcatcg tttcaaagtt 540ggacaacgtc ttgaactatt aaattattcc aattctactg aaatacgcgt agcgcgaatt 600caagaaatat gtggacgacg aatgaatgta tctatcacaa agaaagactt tcccgaatcg 660cttccagatg cagatgacga cagacaagtc tttagctctg gatctcaata ttggatagac 720gagggaagct tcttcatatt tcctgttgga tttgcagcag tcaatggata tcaactaaat 780gcgaaaaagg aatatattga gcacacaaat aaaattgctc aagcaataaa aaatggagaa 840aatccaagat atgactcaga cgacgtcaca tttgatcaat tagcaaaaga tccaattgat 900cccatgattt ggagaaaagt taaggttgga caaaagtttg agctcatcga ccccttggct 960cagcaattca ataacctcca cgtcgcttcg attctcaaat tttgcaaaac tgaaggatat 1020cttattgtgg gaatggatgg tccagatgca cttgaagaca gttttcctat tcatatcaat 1080aatacattta tgttcccagt tggttatgcg gaaaagtata atttggaact tgttccgcca 1140gatgagttca aaggaacatt cagatgggat gaatacttgg agaaagaatc tgcagaaacc 1200ctaccgcttg acttgttcaa gccaatgcct tcctaagaga gattagacaa atttaaggta 1260attctgattt ccaaacgggt aggactacgc cttgaagctg ctgacatgtg tgaaaatcag 1320tttatttgtc cagctacagt gaaatcagtt catggaagac tgataaatgt caatttcgac 1380ggctgggatg aagaatttga tgaactgtat gatgtggact cccatgatat tctaccgata 1440ggatggtgtg aagcgcacag ttatgttcta caacctccga aaaagtacaa ctattga 1497 741497 DNA Caenorhabditis elegans 74 atgtctgaat ttctgaaaat tgtcagagctaacaaaaaat cggacagaaa actcgataag 60 acctacttgt gggaatccta tttacatcagttcgagaaag gaaaaacttc tttcattcca 120 gttgaagcat tcaatcgtaa ccttacagttaattttaacg aatgcgtgaa ggaaggagtt 180 atcttcgaaa cagtggtcca tgattatgacaagaactgcg attcgattca agtcagatgg 240 tttgcacgaa ttgaaaaagt ttgcggatacagagttctgg ctcagtttat cggagctgac 300 acgaaatttt ggctcaatat tttatcggacgatatgtttg gtttggcaaa cgccgcaatg 360 agtgatccca atatggataa aattgtatatgctccgccgc ttgcaatcaa cgaagaatac 420 caaaatgata tggtaaatta tgtaaataattgcattgatg gcgaaatcgt cggccaaact 480 tcgctgtctc caaaattcga tgaagggaaggctctcctaa gcaagcatcg tttcaaagtt 540 ggacaacgtc ttgaactatt aaattattccaattctactg aaatacgcgt agcgcgaatt 600 caagaaatat gtggacgacg aatgaatgtatctatcacaa agaaagactt tcccgaatcg 660 cttccagatg cagatgacga cagacaagtctttagctctg gatctcaata ttggatagac 720 gagggaagct tcttcatatt tcctgttggatttgcagcag tcaatggata tcaactaaat 780 gcgaaaaagg aatatattga gcacacaaataaaattgctc aagcaataaa aaatggagaa 840 aatccaagat atgactcaga cgacgtcacatttgatcaat tagcaaaaga tccaattgat 900 cccatgattt ggagaaaagt taaggttggacaaaagtttg agctcatcga ccccttggct 960 cagcaattca ataacctcca cgtcgcttcgattctcaaat tttgcaaaac tgaaggatat 1020 cttattgtgg gaatggatgg tccagatgcacttgaagaca attttcctat tcatatcaat 1080 aatacattta tgttcccagt tggttatgcggaaaagtata atttggaact tgttccgcca 1140 gatgagttca aaggaacatt cagatgggatgaatacttgg agaaagaatc tgcagaaacc 1200 ctaccgcttg acttgttcaa gccaatgccttcccaagaga gattagacaa atttaaggta 1260 attctgattt ccaaacgggt aggactacgccttgaagctg ctgacatgtg tgaaaatcag 1320 tttatttgtc cagctacagt gaaatcagttcatggaagac tgataaatgt caatttcgac 1380 ggctgggatg aagaatttga tgaactgtatgatgtggact cccatgatat tctaccgata 1440 ggatggtgtg aagcgcacag ttatgttctacaacctccga aaaagtacaa ctattga 1497 75 1497 DNA Caenorhabditis elegans 75atgtctgaat ttctgaaaat tgtcagagct aacaaaaaat cggacagaaa actcgataag 60acctacttgt gggaatccta tttacatcag ttcgagaaag gaaaaacttc tttcattcca 120gttgaagcat tcaatcgtaa ccttacagtt aattttaacg aatgcgtgaa ggaaggagtt 180atcttcgaaa cagtggtcca tgattatgac aagaactgcg attcgattca agtcagatgg 240tttgcacgaa ttgaaaaagt ttgcggatac agagttctgg ctcagtttat cggagctgac 300acgaaatttt ggctcaatat tttatcggac gatatgtttg gtttggcaaa cgccgcaatg 360agtgatccca atatggataa aattgtatat gcttcgccgc ttgcaatcaa cgaagaatac 420caaaatgata tggtaaatta tgtaaataat tgcattgatg gcgaaatcgt cggccaaact 480tcgctgtctc caaaattcga tgaagggaag gctctcctaa gcaagcatcg tttcaaagtt 540ggacaacgtc ttgaactatt aaattattcc aattctactg aaatacgcgt agcgcgaatt 600caagaaatat gtggacgacg aatgaatgta tctatcacaa agaaagactt tcccgaatcg 660cttccagatg cagatgacga cagacaagtc tttagctctg gatctcaata ttggatagac 720gagggaagct tcttcatatt tcctgttgga tttgcagcag tcaatggata tcaactaaat 780gcgaaaaagg aatatattga gcacacaaat aaaattgctc aagcaataaa aaatggagaa 840aatccaagat atgactcaga cgacgtcaca tttgatcaat tagcaaaaga tccaattgat 900cccatgattt ggagaaaagt taaggttgga caaaagtttg agctcatcga ccccttggct 960cagcaattca ataacctcca cgtcgcttcg attctcaaat tttgcaaaac tgaaggatat 1020cttattgtgg gaatggatgg tccagatgca cttgaagaca gttttcctat tcatatcaat 1080aatacattta tgttcccagt tggttatgcg gaaaagtata atttggaact tgttccgcca 1140gatgagttca aaggaacatt cagatgggat gaatacttgg agaaagaatc tgcagaaacc 1200ctaccgcttg acttgttcaa gccaatgcct tcccaagaga gattagacaa atttaaggta 1260attctgattt ccaaacgggt aggactacgc cttgaagctg ctgacatgtg tgaaaatcag 1320tttatttgtc cagctacagt gaaatcagtt catggaagac tgataaatgt caatttcgac 1380ggctgggatg aagaatttga tgaactgtat gatgtggact cccatgatat tctaccgata 1440ggatggtgtg aagcgcacag ttatgttcta caacctccga aaaagtacaa ctattga 1497 762307 DNA Caenorhabditis elegans 76 atgctaaaat tagtcatttt gtgcttcgcgttgttctaca atacagtcag ttcgacaaga 60 tttctgtttg gcgtcgaagt taagtgtgattttgatgaag tgttccaatt aacagtgtcg 120 cattgggaag acgatggcaa tactttttgggatcgcgatg aagacatcac tggacgtatg 180 actatgtttg ctcgaaagaa aatatttttctatcaggacg gccatcatgg atttgaattt 240 ggaaagctcg agccttatgg gtggtttctgcacaattgca cgaaaaatgg aaattttcgc 300 gagtataggc acgggttgag tagcaccagtggatccaatg ggttggagta tattgagtac 360 actgtgaatt tgacgaacgc ctgagaaatatcaaatcaaa tcgaactaac tttcaatttc 420 aataaacatt ctctctaatt acgttttaaaccagtcttaa tttcagatgt ctgaatttct 480 gaaaattgtc agagctaaca aaaaatcggacagaaaactc gataagacct acttgtggga 540 atcctattta catcagttcg agaaaggaaaaacttctttc attccagttg aagcattcaa 600 tcgtaacctt acagttaatt ttaacgaatgcgtgaaggaa ggagttatcg tgagttcata 660 ttgttcgtaa atcggtttta aaatacaatttttgtagttc gaaacagtgg tccatgatta 720 tgacaagaac tgcgattcga ttcaagtcagatggtttgca cgaattgaaa aagtttgcgg 780 atacagagtt ctggctcagt ttatcggagctgacacgaaa ttttggctca atattttatc 840 ggacgatatg tttggtttgg caaagtaagttggacgctca gctctttcta ctattctaaa 900 taaataatgg ttctgttaca taaaattctagagaacaatc gtattaaaac ttcgaaacat 960 ttgtataata gtaaaatttg aacatttcagcgccgcaatg agtgatccca atatggataa 1020 aattgtatat gctccgccgc ttgcaatcaacgaagaatac caaaatgata tggtaaatta 1080 tgtaaatgta agtttgtttt tttccgaatttatgttaata tcatctcaca acttcagaat 1140 tgcattgatg gcgaaatcgt cggccaaacttcgctgtctc caaaattcga tgaagggaag 1200 gctctcctaa gcaagcatcg tttcaaagttggacaacgtc ttgaactatt aaattattcc 1260 aattctactg aaatacgcgt agcgcgaattcaagaaatat gtggacgacg aatgaatgta 1320 tctatcacaa agaaagactt tcccgaatcgcttccagatg cagatgacga cagacaagtc 1380 tttagctctg gatctcaata ttggatagacgagggaagct tcttcatatt tcctgttgga 1440 tttgcagcag tcaatggata tcaactaaatgcgaaaaagg aatatattga gcacacaaat 1500 aaaattgctc aagcaataaa aaatggagaaaatccaagat atgactcaga cgacgtcaca 1560 tttgatcaat tagcaaaaga tccaattgatcccatgattt ggagaaaagt taaggttgga 1620 caaaagtttg agctcatcga ccccttggctcagcaattca ataacctcca cgtcgcttcg 1680 attctcaaat tttgcaaaac tgaaggatatcttattgtgg gaatggatgg tccagatgca 1740 cttgaagaca gttttcctat tcatatcaataatacattta tgttcccagt tggttatgcg 1800 gaaaagtata atttggaact tgttccgccagatgagttca aaggaacatt cagatgggat 1860 gaatacttgg agaaagaatc tgcagaaaccctaccgcttg acttgttcaa gccaatgcct 1920 tcccaagaga gattagacaa atttaaggtaattctgattt ccaaacgggt tgttttatat 1980 cgtttgagat tgtttcacta ttaatagttattcataattg tttcttgttt taaggtagga 2040 ctacgccttg aagctgctga catgtgtgaaaatcagttta tttgtccagc tacagtgaaa 2100 tcagttcatg gaagactgat aaatgtcaatttcgacggct gggatgaaga atttgatgaa 2160 ctgtatgatg tggagtgagt ttatcatgaccgaacgacat tttttcaatg aaaattctat 2220 catttcagct cccatgatat tctaccgataggatggtgtg aagcgcacag ttatgttcta 2280 caacctccga aaaagtacaa ctattga 230777 2307 DNA Caenorhabditis elegans 77 atgctaaaat tagtcatttt gtgcttcgcgttgttctaca atacagtcag ttcgacaaga 60 tttctgtttg gcgtcgaagt taagtgtgattttgatgaag tgttccaatt aacagtgtcg 120 cattgggaag acgatggcaa tactttttgggatcgcgatg aagacatcac tggacgtatg 180 actatgtttg ctcgaaagaa aatatttttctatcaggacg gccatcatgg atttgaattt 240 ggaaagctcg agccttatgg gtggtttctgcacaattgca cgaaaaatgg aaattttcgc 300 gagtataggc acgggttgag tagcaccagtggatccaatg ggttggagta tattgagtac 360 actgtgaatt tgacgaacgc ctgagaaatatcaaatcaaa tcgaactaac tttcaatttc 420 aataaacatt ctctctaatt acgttttaaaccagtcttaa tttcagatgt ctgaatttct 480 gaaaattgtc agagctaaca aaaaatcggacagaaaactc gataagacct acttgtggga 540 atcctattta catcagttcg agaaaggaaaaacttctttc attccagttg aagcattcaa 600 tcgtaacctt acagttaatt ttaacgaatgcgtgaaggaa ggagttatcg tgagttcata 660 ttgttcgtaa atcggtttta aaatacaatttttgtagttc gaaacagtgg tccatgatta 720 tgacaagaac tgcgattcga ttcaagtcagatggtttgca cgaattgaaa aagtttgcgg 780 atacagagtt ctggctcagt ttatcggagctgacacgaaa ttttggctca atattttatc 840 ggacgatatg tttggtttgg caaagtaagttggacgctca gctctttcta ctattctaaa 900 taaataatgg ttctgttaca taaaattctagagaacaatc gtattaaaac ttcgaaacat 960 ttgtataata gtaaaatttg aacatttcagcgccgcaatg agtgatccca atatggataa 1020 aattgtatat gctccgccgc ttgcaatcaacgaagaatac caaaatgata tggtaaatta 1080 tgtaaatgta agtttgtttt tttccgaatttatgttaata tcatctcaca acttcagaat 1140 tgcattgatg gcgaaatcgt cggccaaacttcgctgtctc caaaattcga tgaagggaag 1200 gctctcctaa gcaagcatcg tttcaaagttggacaacgtc ttgaactatt aaattattcc 1260 aattctactg aaatacgcgt agcgcgaattcaagaaatat gtggacgacg aatgaatgta 1320 tctatcacaa agaaagactt tcccgaatcgcttccagatg cagatgacga cagacaagtc 1380 tttagctctg gatctcaata ttggatagacgagggaagct tcttcatatt tcctgttgga 1440 tttgcagcag tcaatggata tcaactaaatgcgaaaaagg aatatattga gcacacaaat 1500 aaaattgctc aagcaataaa aaatggagaaaatccaagat atgactcaga cgacgtcaca 1560 tttgatcaat tagcaaaaga tccaattgatcccatgattt ggagaaaagt taaggttgga 1620 caaaagtttg agctcatcga ccccttggctcagcaattca ataacctcca cgtcgcttcg 1680 attctcaaat tttgcaaaac tgaaggatatcttattgtgg gaatggatgg tccagatgca 1740 cttgaagaca gttttcctat tcatatcaataatacattta tgttcccagt tggttatgcg 1800 gaaaagtata atttggaact tgttccgccagatgagttca aaggaacatt cagatgggat 1860 gaatacttgg agaaagaatc tgcagaaaccctaccgcttg acttgttcaa gccaatgcct 1920 tcccaagaga gattagacaa atttaaggtaattctgattt ccaaacgggt tgttttatat 1980 cgtttgagat tgtttcacta ttaatagttattcataattg tttcttgttt taaggtagga 2040 ctacgccttg aagctgctga catgtgtgaaaatcagttta tttgtccagc tacagtgaaa 2100 tcagttcatg gaagactgat aaatgtcaatttcgacggct gggatgaaga atttgatgaa 2160 ctgtatgatg tggagtgagt ttatcatgaccgaacgacat tttttcaatg aaaattctat 2220 catttcaact cccatgatat tctaccgataggatggtgtg aagcgcacag ttatgttcta 2280 caacctccga aaaagtacaa ctattga 230778 2307 DNA Caenorhabditis elegans 78 atgctaaaat tagtcatttt gtgcttcgcgttgttctaca atacagtcag ttcgacaaga 60 tttctgtttg gcgtcgaagt taagtgtgattttgatgaag tgttccaatt aacagtgtcg 120 cattgggaag acgatggcaa tactttttgggatcgcgatg aagacatcac tggacgtatg 180 actatgtttg ctcgaaagaa aatatttttctatcaggacg gccatcatgg atttgaattt 240 ggaaagctcg agccttatgg gtggtttctgcacaattgca cgaaaaatgg aaattttcgc 300 gagtataggc acgggttgag tagcaccagtggatccaatg ggttggagta tattgagtac 360 actgtgaatt tgacgaacgc ctgagaaatatcaaatcaaa tcgaactaac tttcaatttc 420 aataaacatt ctctctaatt acgttttaaaccagtcttaa tttcagatgt ctgaatttct 480 gaaaattgtc agagctaaca aaaaatcggacagaaaactc gataagacct acttgtggga 540 atcctattta catcagttcg agaaaggaaaaacttctttc attccagttg aagcattcaa 600 tcgtaacctt acagttaatt ttaacgaatgcgtgaaggaa ggagttatcg tgagttcata 660 ttgttcgtaa atcggtttta aaatacaatttttgtagttc gaaacagtgg tccatgatta 720 tgacaagaac tgcgattcga ttcaagtcagatggtttgca cgaattgaaa aagtttgcgg 780 atacagagtt ctggctcagt ttatcggagctgacacgaaa ttttggctca atattttatc 840 ggacgatatg tttggtttgg caaagtaagttggacgctca gctctttcta ctattctaaa 900 taaataatgg ttctgttaca taaaattctagagaacaatc gtattaaaac ttcgaaacat 960 ttgtataata gtaaaatttg aacatttcagcgccgcaatg agtgatccca atatggataa 1020 aattgtatat gctccgccgc ttgcaatcaacgaagaatac caaaatgata tggtaaatta 1080 tgtaaatgta agtttgtttt tttccgaatttatgttaata tcatctcaca acttcaaaat 1140 tgcattgatg gcgaaatcgt cggccaaacttcgctgtctc caaaattcga tgaagggaag 1200 gctctcctaa gcaagcatcg tttcaaagttggacaacgtc ttgaactatt aaattattcc 1260 aattctactg aaatacgcgt agcgcgaattcaagaaatat gtggacgacg aatgaatgta 1320 tctatcacaa agaaagactt tcccgaatcgcttccagatg cagatgacga cagacaagtc 1380 tttagctctg gatctcaata ttggatagacgagggaagct tcttcatatt tcctgttgga 1440 tttgcagcag tcaatggata tcaactaaatgcgaaaaagg aatatattga gcacacaaat 1500 aaaattgctc aagcaataaa aaatggagaaaatccaagat atgactcaga cgacgtcaca 1560 tttgatcaat tagcaaaaga tccaattgatcccatgattt ggagaaaagt taaggttgga 1620 caaaagtttg agctcatcga ccccttggctcagcaattca ataacctcca cgtcgcttcg 1680 attctcaaat tttgcaaaac tgaaggatatcttattgtgg gaatggatgg tccagatgca 1740 cttgaagaca gttttcctat tcatatcaataatacattta tgttcccagt tggttatgcg 1800 gaaaagtata atttggaact tgttccgccagatgagttca aaggaacatt cagatgggat 1860 gaatacttgg agaaagaatc tgcagaaaccctaccgcttg acttgttcaa gccaatgcct 1920 tcccaagaga gattagacaa atttaaggtaattctgattt ccaaacgggt tgttttatat 1980 cgtttgagat tgtttcacta ttaatagttattcataattg tttcttgttt taaggtagga 2040 ctacgccttg aagctgctga catgtgtgaaaatcagttta tttgtccagc tacagtgaaa 2100 tcagttcatg gaagactgat aaatgtcaatttcgacggct gggatgaaga atttgatgaa 2160 ctgtatgatg tggagtgagt ttatcatgaccgaacgacat tttttcaatg aaaattctat 2220 catttcagct cccatgatat tctaccgataggatggtgtg aagcgcacag ttatgttcta 2280 caacctccga aaaagtacaa ctattga 2307

What is claimed is:
 1. A method of identifying a compound that modulatescell proliferation, said method comprising: (a) contacting a cellexpressing a nucleic acid molecule operably linked to a lin-61 promoterwith a candidate compound; and (b) measuring the expression of saidnucleic acid molecule, wherein an alteration in the level of expressionof said nucleic acid molecule, relative to a control, indicates thepresence of a compound that modulates cell proliferation.
 2. The methodof claim 1, wherein said nucleic acid molecule is a lin-61.
 3. Themethod of claim 1, wherein said nucleic acid molecule is a reportergene.
 4. The method of claim 1, wherein step (b) comprises measuring theexpression of the protein encoded by said nucleic acid molecule.
 5. Themethod of claim 4, wherein said protein is contacted with an antibodythat specifically binds to a LIN-61 polypeptide.
 6. The method of claim1, wherein step (b) comprises measuring the mRNA level of said nucleicacid molecule.
 7. A method of identifying a nucleic acid molecule thatmodulates cell proliferation, said method comprising: (a) expressing ina cell (i) a first nucleic acid molecule operably linked to a firstpromoter, wherein said first promoter is a lin-61 promoter; and (ii) asecond nucleic acid molecule operably linked to a second promoter; and(b) measuring the expression of said first nucleic acid molecule,wherein a modulation in said expression of said first nucleic acidmolecule in the presence of said second nucleic acid molecule, relativeto a control, indicates that said second nucleic acid moleculesmodulates cell proliferation.
 8. The method of claim 7, wherein saidfirst nucleic acid molecule is a lin-61 nucleic acid molecule.